#mosquitocidal

[antimnemonic]

being attractive and booby is great. i love when im like "i like bugs actually ^___^" and ppl in the middle of a mosquitocide rant are suddenly like "uwewawewawhuhewah i like bugs too actually!! I'm just scared of roaches/spiders/whatever" extremely funny, happens all the time

[aistobascistod]

Mosquitocide : No more mosquitos :: Coincide : No more coins

[medtalksblog]

Reproductive Function Impairment in Men and the Potential of Tridax Procumbens

Reproductive function issues in men are a prevalent medical condition that can lead to problems like infertility, reduced libido, and erectile dysfunction. These problems significantly impact sexual functioning and overall health. Hormonal imbalances, physical abnormalities, and lifestyle choices, such as smoking and excessive alcohol consumption, can contribute to these impairments. Fortunately, there are various treatments available to help men manage and enhance their reproductive function.

Studies by researchers have confirmed that the aqueous leaf extract of Tridax procumbens can mitigate reproductive function impairments. The extract improves sperm quality, reduces oxidative stress, balances hormones, restores healthy glucose levels, and enhances sexual performance when taken orally.

Tridax procumbens, a wild plant in the Asteraceae family, found in tropical and subtropical areas, especially in India, has been used in traditional Ayurvedic medicine for years. Extracts from its leaves, stem, flower, and roots are rich in alkaloids, steroids, carotenoids, flavonoids, fatty acids, phytosterols, tannins, and minerals, used to treat diabetes, arthritis, and inflammatory reactions. Experimental studies have demonstrated its antioxidant, antibacterial, anti-inflammatory, antimicrobial, and mosquitocidal activities.

Currently, the aqueous leaf extract of Tridax procumbens is gaining attention for its potential benefits on reproductive health and sexual performance in men. It's becoming popular as an Ayurvedic treatment for infertility, weak erection, erectile dysfunction, and premature ejaculation.

From a clinical perspective, a healthy erection requires proper blood circulation, suitable hormone levels, a functional nervous system, and a healthy psychological outlook. These components are often compromised in individuals with long-term medical issues like diabetes, hypertension, abnormal cholesterol levels, and obesity.

Tridax Procumbens has various positive effects on the body, including vasodilation and improved blood circulation, aiding in penile erection. It relaxes blood vessels, enhances blood flow to the penis, and strengthens an erection, assisting in the treatment of erectile dysfunction. Additionally, it boosts sperm production and increases serum testosterone concentration, improving male sexual performance and sex drive. In an experimental study, Tridax Procumbens was found to have effects on male fertility similar to Viagra.

[mosquitopunks-blog]

PREMIUM HIGH QUALITY AND ALL-NATURAL INGREDIENTS: Typhaceae fiber, Makko powder, Bamboo stick, Pine resin, Tragacanth gum and your favorite aroma ENTIRELY DEET FREE: Our Mosquitocide incense sticks are made entirely free of DEET BURN TIME: Each stick burns up to 1 - 1 1/2 hours. Each Tube of MosquitoPunks provides 20 h

[gopeshsblog]

Tridax procumbens, commonly known as coatbuttons or tridax daisy, is a species of flowering plant in the daisy family. It is best known as a widespread weed and pest plant. It is native to the tropical Americas, but it has been introduced to tropical, subtropical, and mild temperate regions worldwide.  It's used majorly in Indian traditional medicine. This is rich in alkaloids, steroids, carotenoids, flavonoids (such as catechins, centaurein and bergenins), fatty acids, phytosterols, tannins and minerals. Concoctions of extracts from T. procumbens leaves, stem, flower, and roots are used to treat patients suffering from diabetes, arthritis, inflammatory reactions and even applied to open wounds. The medicinal value of extracts has been evident by in vitro/in vivo assay of antioxidant, anti-bacterial, anti-inflammatory, anti-microbial, vasorelaxant, anti-leishmanial and mosquitocidal activities. Still, there is derth in the studies on isolation, characterization and evaluation of active principles from the extracts. This current review article gives comprehensive information about the T. procumbens taxonomy, morphology, geographical distribution, phytoconstituents and pharmacological activities. #Gopesh @Gopesh. #tridax_procumbens #tridax_daisy #floral #flowers_mania__ #flowers #flowerstagram #flowerphotography #flowers_beauties #flowersofinstagram #flower_shotz #flowers_on_insta #flower_perfection #flowers🌸 #nature #nature_shot #nature_lovers #naturephotography #nature_brillance #nature_perfection #nature_beautiful #nature_beauty #nature_photography #nature_of_our_world #nature_brilliance #naturelovers #macro_admirer #macromania #macro_perfection #macro_ https://www.instagram.com/p/CI_B8bysOhS/?igshid=1ry53wh9uzsfj

[apexherbals]

Curry Leaves

Karapincha or Kadi Patta is that the leaves of the curry tree and are scientifically referred to as Murraya koenigii sprang. It belongs to the Rutaceae family. The plant is endemic to the Republic of India and is often found within the tropics and semi tropics. it's cultivated in numerous countries as well as China, Australia, Nigeria, and country. The height of the plant is six to fifteen feet. The helpful components of this plant area unit its leaves, roots, and bark. The leaves have invariably been looked at for their distinctive flavor and cooking edges, however, they even have a variety of spectacular health edges. looking at the supposed use, the leaves will dry or fry. The recent type is additionally highly regarded for preparation and seasoner drugs. The leaves area unit employed in numerous native cuisines throughout the Republic of India and Asia with nice seasoner properties. Curry leaves area unit kind of like the Indian lilac or Indian lilac, and in several Indian languages,​​their name is translated as sweet arishth. Curry Leaves Nutrition Facts The main nutrients found in curry leaves area unit carbohydrates, fiber, calcium, chemical element, iron, magnesium, copper, and alternative minerals. They additionally contain numerous vitamins like B3 (nicotinic acid) vitamin C, vitamin A, vitamin B, vitamin E, antioxidants, plant sterols, amino acids, glycosides, and flavonoids. They even have a trace, nonsignificant quantity of fat (0.1 g per one00 g). Another chemical constituent gift in curry leaves is carbazole alkaloids. a quest study revealed within the Journal of Agricultural and Food Chemistry showed that alkaloids found within the leaves possess inhibitor properties. Carbazole alkaloids embody mahanimbine, murrayanol, mahanineoenimbine, O-methylmurrayamine A, O-methylmahanine, isomahanine, bismahanine and bispyrayafoline. an additional study conducted at the Department of gardening at Michigan State University recommended that these chemicals have insecticidal and antimicrobial properties additionally, specifically mosquitocidal properties Benefits of Curry Leaves Most people assume that curry leaves area unit solely supplementary for flavor and throw the leaves away whereas intake their soup or curry. However, they're way more vital than several understand, and provide a variety of health edges with none aspect effects.

https://www.etsy.com/listing/927608383/organic-curry-leaves-and-powdermurraya?ref=shop_home_feat_2&frs=1

[soumyamajumder]

GC-MS analysis reveals Dendrobium candidum is a mosquito-attractant orchid with mosquitocidal compounds. Full article is now on researchgate.

[montclarelab]

A Tryst with Science

By Ashwitha Lakshmi 

Here I am, sipping my coffee and staring out of my only real window to the outside world. It all does seem a bit too cinematic. And as a clockwork, I start to ponder on the endless rhetorical questions that we often ask ourselves. 

Isn’t it amusing how we are so caught up in the present (or future) that we lose track of how far we have come? A typical Indian twenty-something, in New York, living a life she still finds hard to believe. A student in a renowned University and an even amusing Laboratory. I get to do everything that I ever dreamt of, and more. Can life get any better?

I can see my reflection smiling away in the window and I think “Oh I know what comes next”. I’m pulled into a known reverie.

As cliche as it sounds, unlike most, I do not remember when or how I fell in love with science. But one memory does come to my mind. In 2007 I got to meet my grandparents after a long hiatus (isn’t that always the case?) but they were very ill and I felt utterly helpless. For a brief period of time, I wanted to become a doctor and help others who were sick. But to my dismay, I was made aware that even the doctors often felt helpless. 

And that is when a realisation hit me - the medical profession stands tall on the shoulders of thousands of scientist’s life work. And for doctors to function, we need scientists who lock themselves in labs for years together, if not decades. (More power to doctors, especially in tempestuous times like these, but we as scientists have a hugley pivotal role to play)

My first rendezvous with anything remotely “sci-fi” was when I met my uncle and aunt in their lab. Yes, they are scientists. My uncle was working on this curious project on Kala azar (also known as Visceral Leishmaniasis) which is the second largest parasite killer in the world, which I could hardly follow back then. But it did strengthen my resolve to become a scientist.

Even at the mere mention of a famous scientist’s name, I’d ask myself: What does it take to become a good scientist? And do I have what it takes? What makes me fit for this fascinating but immensely challenging life-career? 

“I am a good human first, meticulous next, then a team player. A contribution made in the spirit of humanitarianism - is what makes a good scientist” - I would pride myself into thinking. 

When I entered Grad school I was worried that I’d spend my entire life with Biology. Thankfully Biotech turned out to be an interdisciplinary field. (*Pheww) And like every grad student, there were times when I thought I had bitten more than I could chew. I would often wonder why I was made to study Engg mechanics and Engg drawing. And there were times when I felt out of place. Call it frustration or just wishful thinking, I even lured myself into thinking that I could become an actor instead. But life has weird ways, even a small mosquito can induce life changing decisions (intended cliff hanger).

As weird as it may sound, a sleepless (blame it on mosquitoes) night can actually change the course of your life. Weirdly, my research idea on mosquito proteins gave me an opportunity of a lifetime - to work in one of the most stellar labs in India - BARC (Bhabha Atomic Research center) where I worked in Radiation biology and health science department on Expression and purification of mosquitocidal binary toxin component BinB and its receptor protein Cqm1 using bacterial expression system project. For me, all this while science was just in the books or in the research papers. But for the first time, I could hold a protein in my hands (Of course immobilized in gel ;) ). The first time I could visualize a pure protein band - a huge one at that! 

All of these were baby steps to me in the field of scientific research and I couldn’t wait for more. 

My usual lab schedule was between 8 in the morning till 8 in the evening and I still yearned for more (and nope, I’m not bragging ;) ). Nobody could stop me from thinking about going back to the lab every day. On some days I couldn’t even sleep due to excitement. The next seven months went in a jiffy. And all the hard work culminated into a Research Publication. 

When I look back it was one of the greatest achievements of my life. From begrudgingly referring to research papers for class assignments, to writing and publishing my own research paper with “actual scientists”, it has been an exhilarating journey.

That’s when I knew I wanted to spend my life in scientific research. The new-found purpose of my life made me pursue a Masters degree in Biotechnology. And I remember being overjoyed to the extent of crying, when I was accepted into NYU. Once in post-grad, I worked as a Graduate teacher’s assistant for the first 6 months and then received an email from my advisor on an opening for graduate Research assistant in Montclare lab and after reading the work that was happening there on protein, I knew I had to apply. 

The first formal job interview of my life! The excitement was killing me but it helped me prepare for it. On the day of the interview I remember being scared out of my life but I gathered all my courage and reached for the interview where I met Stanley Chu (my mentor). He put me in ease with his demeanour and kind words. And the questions were pretty interesting too. Overall, I was happy with the way the interview turned out. (Fingers crossed)

I was in my class when I received an email saying I got the job and cried from happiness again (I cry a lot when I’m happy lol). Next thing I know I am on a video call with my whole family and friends, and was super excited to begin work at this lab. In hindsight, I could not have asked for a better mentor and was a bit too excited to learn alot from him.

Life has been kind to me with wonderful people - my parents, my elder sister Pavi, all my friends at high school and grad school. Yes, good people are often hard to come by, much harder to retain. But I am lucky in that sense because I have my people whom I can trust my life with. Oh the gratitude and compassion! I feel a new spring of warmth blossoming inside of me. 

Now I see my reflection smiling away in the window and now I know why. And I’m pulled back into reality.

[thehollow06-blog]

Industrial applications of keratin: more than feathers

Pablo Alarcón

00136791

Executive Summary

Poultry industry is increasing hand in hand with population increase. The reason being that chicken meat is the most consumed by humans. However, not only the meat is consumed. People can use chicken feathers for different industrial applications, and this practice is environmentally friendly, because feathers are difficult to degrade. The industrial applications are a feasible way to not throw away feathers, and it’s because the main component of chicken feathers is keratin which is a protein that has sulfur in its molecular structure. Many products have been elaborated with keratin from chicken feathers such as biofilms, fibers, shampoos and so on.

Introduction

The population's demand for food has increased, seeking that food security is met for the entire population, and one consequence of this problematic is the big progress of poultry industry. The production of chicken meat was about 90-100 million of tons, being the most producing countries Brazil and United States (Mothé, Viana & Mothé, 2017). Of course, the residues also increase, so different industries could use it in determined applications since it is microbiologically allowed. This is considered as an alternative instead of burying or burning chicken feathers. For use the main component of feathers, it has to take place many chemical reactions, so then keratin could be used. It’s important to know that keratin is a complex compound, so it is complicated to hydrolyze it (Chandrakant et al., 2017). In the middle of the past century, the first publication about an application of chicken feathers mentioned that this is a material that could be used as a fiber. So it is a good option for non-comestible applications. Nevertheless, it could be used as a source of protein for many domestic animals, combined with other nutrients.

Timeline of related investigations

In this century, the first investigation about the extraction of keratin was presented by Coello et al. (2003). In the study, scientists extracted the keratin using the degradation of soil microorganisms, specifically, the bacteria Kokuria rosea. The application of this study was using the extracted keratin in Food supplements for salmons. Feeding salmons with this kind of supplement, made them more colorful. Subsequently, Sayed et al. (2005) took a big step in chicken feathers applications. This group proposed that extracted keratin could be used in polluted water treatment. Specifically, keratin could be used as a bioremediatory compound for different pollutants in this type of water such as calcium, iron, magnesium and so on. Then, Poopathi & Abidha (2007) presented a microbiologically important study. They obtained keratin from chicken feathers and used it in a microbiologically medium, for the bacterial growth. This is a successful application because for each study, laboratory workers need a lot a medium, and it is better for the environment if one of the products of the medium is reused. Also, they used different bacteria specie for de hydrolysis of keratin. After the first decade of this century, Aguayo et al. (2011) followed the line of investigation of Sayed et al. (2005). Also, they presented a study that showed the use of keratin as a component in bioremediation process. In this study the range of metal contaminants was improved.

The food industry was beneficiated with the study of Darah et al. (2013), because they identified another way to use keratin of chicken feather as a food supplement. The presentation in this case was a feather flour. The difference between this study from the study of Coello et al. (2003), is the use of another microorganism specie such us Microsporum fulvum. In the next year, two important studies took place. Xu & Yang (2014), inspired in the first applications of keratin from chicken feathers. They made a fiber with this main component and did physical and chemical analysis for determine the spinning capacity of this fiber. Marculescu & Stan (2014), took another big step in this field. They proved the capacity and the yield of the extracted keratin as a biofuel. However, it showed that is better than a biofuel from wood, but it had a little content of carbon, which is needed to produce CO2. Then, many studies took place. Kawara et al. (2015) used a thermal treatment for produce activated carbon. This study was economically successful because it reduced the cost of produce activated carbon which is used for cosmetical products and for mediums used in in vitro culture of plants. On the other hand, Reddy (2015) made biofilms with this component with similar properties to its counterpart. In the same line of investigation, Poole & Church (2015), made more resistant biofilms so they can be an environmentally friendly alternative to plastics such as HDPE, PA6 and so on.

Another gold year for this type of investigations was 2017. Sharma et al (2017) evaluated the possibility of using the extracted keratin in pharmacological applications such as shampoos and creams. For this application, a chemical method of extraction was the best option. Mothé et al. (2017) followed this line of investigation and improved shampoos based on the extracted keratin. The hairs proved were softer and without frizz. On the other hand, Esparza et al. (2017), made a good example of plastic based on this product. The process was economic, and the physical properties were similar to their counterpart. Finally, Garrido et a. (2018) made biofilms based on soy and the extracted keratin. Also, the process of extraction was economic because it was similar to the other chemical process.

Classification of keratin applications

In summary, extracted keratin from chicken feathers have many fields of applications. In agroindustry applications can be considered the study of the NYSM medium of Poopathi & Abidha (2007), and the study of Reddy (2015) to make biofertilizers. In the cosmetic industry can be considered the study of Sharma et al. (2017), for make shampoos and creams based on this keratin. Mothé et al. (2017) also incurred in this investigation and improved the characteristics of the conventional shampoos. Keratin as a film and fiber were presented by Reedy (2015), Poole & Church (2015), Esparza et al. (2017), Garrido et al. (2018), Xu & Yang (2014). Furthermore, keratin from chicken feathers presented as a main component for food supplements in the studies of Coello et al. (2013).  Finally, this compound proved in bioremediation process by Aguayo et al. (2011).

Conclusions

Keratin from chicken feathers can be used in many industries, and it could be extracted with chemical and biological methods. There aren´t too many studies on this topic, so it has potential to be studied. The most complicated challenge is to see which way to extract keratin is the most economical, most environmentally friendly and in turn reproducible anywhere in the world.

References

Ismael Alejandro Aguayo-Villarreal, Adrián Bonilla-Petriciolet, Virginia Hernández-Montoya, Miguel A. Montes-Morán, Hilda E. Reynel-Avila, Batch and column studies of Zn2+ removal from aqueous solution using chicken feathers as sorbents, In Chemical Engineering Journal, Volume 167, Issue 1, 2011, Pages 67-76, ISSN 1385-8947, https://doi.org/10.1016/j.cej.2010.11.107.

Coello, N., Bernal, C., Bertsch, A., Estrada, O., Moccó, Y., & Hasegawa, M. (2003). Feathers as agro industrial waste: Their biotechnological utilization to develop new added value products. [Las plumas como residuo agroindustrial: Su utilización biotecnológica para producir insumos de interés industrial] Revista De La Facultad De Ingenieria, 18(3), 119-126.

Darah, I., Nur-Diyana, A., Nurul-Husna, S., Jain, K., & Lim, S. -. (2013). Microsporum fulvum IBRL SD3: As novel isolate for chicken feathers degradation. Applied Biochemistry and Biotechnology, 171(7), 1900-1910. doi:10.1007/s12010-013-0496-4

Garrido, T., Leceta, I., de la Caba, K., & Guerrero, P. (2018). Chicken feathers as a natural source of sulphur to develop sustainable protein films with enhanced properties. International Journal of Biological Macromolecules, 106, 523-531. doi:10.1016/j.ijbiomac.2017.08.043

Esparza, Y., Ullah, A., & Wu, J. (2017). Preparation and characterization of graphite oxide nano-reinforced biocomposites from chicken feather keratin. Journal of Chemical Technology and Biotechnology, 92(8), 2023-2031. doi:10.1002/jctb.5196

Marculescu, C., Stan, C. (2914) Pyrolysis treatment of poultry processing industry waste for energy potential recovery as quality derived fuels, In Fuel, Volume 116, Pages 588-594, ISSN 0016-2361, https://doi.org/10.1016/j.fuel.2013.08.039.

Mothé, M. G., Viana, L. M., & Mothé, C. G. (2017). Thermal property study of keratin from industrial residue by extraction, processing and application. Journal of Thermal Analysis and Calorimetry, 1-10. doi:10.1007/s10973-017-6845-8

Poole, A. J., & Church, J. S. (2015). The effects of physical and chemical treatments on Na2S produced feather keratin films. International Journal of Biological Macromolecules, 73, 99-108. doi:10.1016/j.ijbiomac.2014.11.003

Poopathi, S., & Abidha, S. (2007). Use of feather-based culture media for the production of mosquitocidal bacteria. Biological Control, 43(1), 49-55. doi:10.1016/j.biocontrol.2007.04.019

Reddy, N. (2015). Non-food industrial applications of poultry feathers. Waste Management, 45 doi:10.1016/j.wasman.2015.05.023

Xu, H., & Yang, Y. (2014). Controlled de-cross-linking and disentanglement of feather keratin for fiber preparation via a novel process. ACS Sustainable Chemistry and Engineering, 2(6), 1404-1410. doi:10.1021/sc400461d

[juniperpublishersmicrobiology]

Variable Region of 16s rRNA is Essential For the Identification of Group 1 Mosquito-Pathogenic Strains of Lysinibacillus

Authored by  Ashwani Kumar

Abstract

A study was carried out to characterize and identify the potential mosquito-pathogenic bacilli strains isolated from the soil samples obtained from various locations in Goa, India. Six isolates were characterized morphologically, biochemically and phylogentically. Phylogenetic relationships of these strains were determined based on comparison of the 16S rRNA sequences with that of the closest Lysinibacillus species obtained from nucleotide database. Their mosquitocidal abilities were confirmed by bioassay and presence of genes responsible for pathogenicity and toxin production. Molecular characterization by partial sequencing of 16S rRNA gene has confirmed their identity as Lysinibacillus sp. Group 1.

Comparison of the 16S rRNA sequences with that of the five groups of "Bacillus sphaericus like organisms”, re-designated off lately as Lysinibacillus, revealed that a variable region of 695 bp and hypervariable region of 22 bp were highly conserved among these five groups. These isolates were found effective against Culex quinquefasciatus larvae with LC50 values ranging from 0.018 to 0.58 ppm. The PCR amplification also showed the presence of binA, binB binary genes and mtx1, mtx2, mtx3 mosquitocidal genes. Larvicidal activity against Culex quinquefasciatus appeared related with expression of binary toxins. These strains were found more potent than the commercial strain and hence could be formulated for controlling Culex species transmitters of Japanese encephalitis and filariasis.

Keywords: Biocontrol agent; Binary toxins; Hypervariable; Lysinibacillus strains; 16S rRNA gene

Abbreviations: Btx: Binary Toxins; Mtx: Mosquitocidal Toxins; HV: Hyper Variable; NJ: Neighbor Joining; NYSM: Nutrient Yeast Sporulating Medium

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Introduction

Some strains of "Bacillus sphaericus like organisms” are toxic to dipteran larvae and used operationally for control of mosquito-borne diseases. These micro-organisms are aerobic, spore-forming bacteria producing spherical or oval endospores and contain endotoxins responsible for the killing of mosquito larvae [1] The larvicidal strains contain two types of insecticidal toxins viz. binary and mosquitocidal. The binary toxins (Btx) are spore associated toxins produced during sporulation phase and form crystals in the mother cell. The crystal comprises of two toxic proteins with molecular weights of about 42 kDa (binA) and 51 kDa (binB). Both proteins are required in equal amounts for larval toxicity [2,3].

In contrast, mosquitocidal toxins (Mtx) are produced during vegetative stage. There are three Mtx toxins (Mtx1, Mtx2 and Mtx3) that have been identified in some mosquitocidal strains [4-6]. Interestingly, Mtx proteins are degraded by extracellular proteases which are produced when the cell growth reaches sporulation phase [7 ]. Highly toxic strains are known to produce both Btx and Mtx toxins, while other strains synthesize either of them [8]. Krych et al. [9] on the basis of DNA-DNA homology demonstrated that Bacillus sphaericus is a conglomerate of 5 distinct homology groups. Group I represents Bacillus sphaericus sensu stricto, while Group II consists of two subgroups, IIA & IIB. All mosquito pathogenic strains are placed in subgroup IIA. Group IIB consists of non-pathogenic Bacillus fusiformis.

Alexander & Priest [10] attempted to use phenotypic features to separate these groups. The toxic strains of Group IIA produce ribosomal RNA gene restriction patterns which are different from Group IIB and non-toxic Bacillussphaericus sensu stricto [11]. De muro & Priest [12] eveloped an oligonucleotide probe based on specific region of 16S rDNA which differentiates Group IIA from other DNA homology groups. However, Jhanz et al. [13] showed that this probe also hybridizes with non-toxic strains. RAPD and repetitive primer amplified polymorphic DNA analysis showed close relatedness among mosquito pathogenic strains belonging to homology group IIA [14,15]. Nakamura [16] redistributed Bacillus sphaericus-like organism based on 16S rRNA gene sequence into seven distinct clusters. In this schema, mosquitocidal and few non-mosquitocidal strains were clustered in Group 1 which corresponded to Krych Group IIA and closely linked to Group 2 representing Krych Group IIB named as Bacillus fusiformis.

Group 1 to 4 of Bacillus sphaericus like organisms were re-designated as new genus Lysinibacillus on the basis of the presence of lysine in the cell wall [17]. In the past, different techniques have been used for grouping and classifying mosquito pathogens. Except the pathogenicity to mosquito larvae, there are no definitive characters that can separate this from other taxa, therefore the taxonomic status of this group is still unclear.

Here, we have studied variable and hypervariable regions of 16S rRNA gene specific to the strains of Lysinibacillus which correspond to group 1 to 5 of Nakamura [16]. This study was aimed at finding whether the distinctive phenotypic characters, the presence of unique nucleotide sequences and mosquitocidal genes altogether could form basis for distinguishing this group from other spore forming Lysinibacillus.

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Materials and Methods

Source of strain

The bacterial strains were isolated from soil samples which were collected from various mosquito breeding habitats viz., stagnant ponds, paddy fields and mangrove vegetations at different location in Goa, India [18]. Stock cultures were maintained on NYSM agar slants stored at 4 °C.

Characterization of the isolates

Morphological, biochemical and physiological characteristics of the isolates were studied as per Krych et al. [9] and Alexander & Priest [10].

Amplification of 16S rRNA gene of the isolates

The genomic DNA was extracted from 24hrs old culture grown in J-Broth using the method described in Sambrook et al. [19]. Gene of 16S rRNA was amplified using the universal primers, a forward primer (S-D-Bact-0011-a-5-17: 5'-GTTTGATCCTGGCTCAG-3') and a reverse primer (S-*-Univ- 1392-b-A-15: 5'-ACGGGCGGTGTGTNC-3'). The conditions for PCR amplification were used as described by Prabhu et al. [20]. PCR products were purified using purification kit (Quiagen). Purified PCR product obtained was outsourced to Bangalore Genei for sequencing. The sequences obtained were submitted in the Gene Bank under the accession numbers FJ473362 (KSD- 1), FJ473363 (KSD-2), FJ473364 (KSD-3), FJ473365 (KSD-4), FJ473368 (KSD-8) and FJ473369 (KSD-7).

The sequences obtained were compared with those available in NCBI database using BLASTn. The sequences from phylogenetic groups described by Nakamura [16] for Bacillus sphaericus like organisms were compared. Clustal X version 2.0.7 was used to generate multiple sequence alignment between these selected sequences. Consensus sequences obtained from these multiple alignment within groups were compared with sequences available in NCBI databases. Consensus sequences for these groups were then analyzed in relation to variable and hypervariable (HV) regions to develop phylogenetic tree using neighbor joining (NJ) method. Tree was obtained with 1000 seeds and 10000 bootstraps. Final tree was rooted and drawn using MEGA4 [21].

PCR amplification of genes encoding the mosquitocidal toxins

Genomic DNA of isolates was subjected to PCR amplification to detect the presence of genes encoding toxins. Primers used for amplification of toxin genes binA, binB, mtx1, mtx2 and mtx3 are listed in Table 1. Conditions used for amplification were as described in [6,22,23].

Source of immature of test mosquito species

Culex quinquefasciatus larvae were obtained from the insectary of National Institute of Malaria Research, Field Unit, Goa, where this mosquito species is maintained at a temperature of 27±2 °C, relative humidity of 70±5% and a photoperiod: scotoperiod of 12:12 h (light: dark). From this laboratory bred mosquitoes, 3rd instars larvae were selected and used for larval bioassays to determine larvicidal activity of bacterial strains.

Preparation of Lyophilized powder for Bioassay

Isolates were grown in a 250ml conical flask containing 100ml of NYSM broth and incubated at 30 °C on a rotary shaker for 96 hrs. Spore crystals were harvested by centrifugation at 5000rpm for 10 min, cell pellet was washed twice with sterilized distilled water and the final pellet was kept for lyophilization which was carried out in Delvac Model (Lyodel) at temperature -40 °C. Vacuum was applied for 4-6 hours for complete drying. Lyophilized culture was stored at 4 °C until use.

Larval Bioassays

Stock dilutions were prepared by dissolving lyophilized powder in sterile distilled water. Dilutions were made to obtain appropriate range of dosage [24]. For each of the doses, 3 replicates of 25 healthy III instar larvae of Culex quinquefasciatus were introduced in 250ml plastic bowl containing 100ml of sterile distilled water. Concurrent control was maintained under similar conditions without the addition of spore suspension. The mortality was recorded by counting live larvae after 24 & 48 hours. LC50 were determined by Probit analysis with SPSS PASW 18.0 indicating mean and standard errors [25].

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Results and Discussion

Phenotypic characterisation of isolates

Colonies of all six mosquito pathogenic isolates were white- creamish in colour. The vegetative cells were Gram's positive, long and rod-shaped. Sporulating cells consisted of racket shaped swollen sporangia were containing round endospores associated with the crystal proteins. These isolates did not produce nitrate reductase, amylases, tyrosinase and showed the production of catalase, urease and acid from glucose. All isolates grew in broth with 2 to 7% NaCl, sensitive to erythromycin and resistant to streptomycin. These isolates cleared adenine and were unable to utilize citrate and L-isoleucine in the medium.

Earlier, mosquito-pathogenic bacterial strains were classified on the basis of limited biochemical tests as Bacillus sphaericus Krych et al. [9] demonstrated using DNA-DNA homology that Bacillus sphaericus actually a conglomerate of 5 homology groups. The mosquito-pathogenic strains belong to subgroup IIA. Non-pathogenic strains belong to subgroup IIB were identified as Bacillus fusiformis.Alexander & Priest [10] used several phenotypic tests in order to give features specific to mosquito-pathogenic strains belong to subgroup IIA. The above isolates showed all the features in agreement to the described feature of subgroup IIA. It was noted that subgroup IIA pathogens separated from the subgroup IIB non-pathogens only by the ability of the former to clear adenine from nutrient agar and sensitivity to erythromycin. An additional feature is mosquito-pathogenicity, which is based on the production of toxins. Recently, members of group I to IV of Krych et al. [9] were re-designated as members of genus Lysinibacillus due to presence of lysine in the cell wall [17].

Phylogenetic characterization

Sequences of 16S rRNA genes obtained from the isolates were used to search NCBI databases using BLASTn which showed high similarity with Lysinibacillus fusiformis and Lysinibacillus sphaericus. (Figure 1) presents phylogenetic tree based on comparison of the variable region of 16S rRNA sequences generated with that of sequences of species belonging to Lysinibacillus genus, including strains of Bacillus sphaericus like organisms earlier reported as pathogenic to mosquitoes in the GenBank database. Phylogenetic tree separates these organisms into five groups. All the six mosquito-pathogenic isolates were clustered in Group 1 in accordance with Nakamura [16] that correspond to sub-group IIA as described above.

Consensus sequences derived in this study for five groups of Lysinibacillus are shown in (Figure 2). A variable region (V) of 695 bp nucleotide position stretching from 62 to 756 bps and a hypervariable (HV) region of 22 bps at nucleotide position from 178 to 199 bps were found among all these five groups. The HV regions were conserved and specific to respective groups. Interestingly, the highly conserved HV region in Group 1 and Group 2 were different only by three nucleotides at position 197199 bps. Additionally, we found six single nucleotide differences in the variable regions of Group 1 and 2 which were used to differentiate members of these groups. These unique nucleotide differences among these two groups are at nucleotide position 85, 221, 571, 586, 665 and 720.

Group 2 and Group 3 of 16S rRNA gene alignments correspond to sub-group IIB and Group I of DNA-DNA homology and were named as Lysinibacillus fusiformis and Lysinibacillus sphaericus, respectively. Group 4 has been named as Lysinibacillus boronitolerans which corresponds to group III of DNA-DNA homology [26]. So far Group 1 containing mosquito pathogenic strains and Group 5 (DNA-DNA homology Group IV) has not been recognised as a separate species.

The phylogenetic tree shown in (Figure 1) included the consensus sequences drawn for five groups. Sequence similarities of the above mentioned variable region between these five groups are shown in (Table 2). This analysis reveals that Group 1 and Group 2 have maximum commonalities of only 98%. Our findings showed that variable region was sufficient in distinguishing the five groups as the nodes were showing very high bootstrap values (Figure 1). These findings match with the phylogenetic tree drawn using 16S rDNA sequences more than 1400 bases [16]. Although, HV regions showed the signature sequences of Group 1 and Group 2 but were incapable of separating these groups on its own during phylogenetic analysis. This region was earlier used by de Muro et al. [12] for designing probe for identifying the mosquito-pathogenic isolates.

Mosquito-pathogenicity of isolates

+, positive; -, negative; V, variable;

*Data from other study as follows: Group2, Group3, Group4 and Group5 organism's biochemical characteristics as described in Alexander& Priest [10].

All 6 isolates were tested against laboratory reared III instar larvae of Culex quinquefasciatus. These isolates were found highly toxic to Culex quinquefasciatus larvae, with LC50 values ranging from 0.05to 0.87 ppm at 24 hrs post treatment and 0.018 to 0.58 ppm at 48 hrs (Table 3). Isolate KSD-2 had lowest LC50 value of 0.05 & 0.018 ppm at 24 and 48 hrs respectively and hence was most lethal. Lethal conc. of KSD-2 was 32 folds lower as compared with commercial Bacillus sphaericus 101 (H5a5b)

PCR amplification of binary and mosquitocidal genes confirmed the presence of binA, binB, mtx1, mtx2 and mtx3 in all 6 isolates as shown in (Figure 3). Larvicidal activity was found to be primarily associated with binary toxins. This could be explained as only sporulating stage was tested for determination of larvicidal activity. Previous studies also showed that highly toxic strains are known to produce both binary and Mtx toxin proteins, while other strains synthesize either of these toxins [8]. The DNA sequences of the genes encoding these proteins are highly homologous between the strains, although they were isolated from different continents [27]. Previously, pathogenicity of Bacilli was considered important for distinguishing taxa as there were no reliable phenotypic characters [10], but now probe based on 16S rRNA and toxin genes are available to segregate mosquito-pathogenic strains morphologically similar to Bacillus sphaericus like organisms [12,28]. In the present study, the presence of both bin and mtx genes in the mosquito- pathogenic strains has been confirmed.

Proposal for species novo

Classical characterization methods for describing Bacillus sphaericus have failed to provide enough taxonomic information [29]. Krych et al. [9] were the first to identify 5 distinct DNA-DNA homology groups among the 50 strains, which include 7 strains showing pathogenicity to mosquito larvae, which were placed in the sub-Group IIA. There was about 62% DNA-DNA sequence homology, with ATm of about 7oC between the strains of subgroup IIA and IIB. It is generally argued that strains within a species show greater than 70% DNA sequence homology and thermal hybrid stability (ATm) of less than 6oC [30]. This amply suggested that sub-group IIA and IIB cannot be allocated to the same species. The creation of a separate species for the mosquito pathogens was contemplated earlier but due to lack of positive phenotypic tests, other than mosquito pathogenicity, its identification as a taxon has been delayed. Alexander &Priest [10] used several phenotypic tests and tried to develop numerical taxonomic group in order to identify features specific to mosquito-pathogenic strains belonging to subgroup IIA. Further support for distinction of sub-group IIA pathogens from non-pathogenic sub-group IIB became possible by rRNA gene pattern [11], RAPD finger print and isoenzyme analysis [31].

These studies revealed a very low similarity between the two subgroups and led to the conclusion that the pathogenic strains are an independent lineage distinct from the other round spore forming bacilli of sub-group IIB. Nakamura [16] attempted to raise the status of sub-group IIA and sub-group IIB to independent groups namely Group 1 and Group 2, respectively using 16S rRNA sequences. Now with re-designation of the genus due to presence of Lysine in cell wall the sub-group IIB is known as Lysinibacillus fusiformis.

PROBIT model: PROBIT(p) = Intercept + BX (Covariates X (dose) are transformed using the base 10.000 logarithm)

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Acknowledgement

We thank the Indian Council of Medical Research, National Institute of Malaria Research, New Delhi for institutional support. We are thankful to the field and laboratory staff of National Institute of Malaria Research (ICMR), Field Unit, Goa, India. The authors also thank Dr. Barbuddhe of ICAR, Research Complex, Goa for providing the Lyophiliser facility. This paper bears the NIMR publication screening committee approval no. 049/2016.

The present investigation clearly identified the differences of 16S rRNA sequences between various members of Lysininbacillus including Lysinibacillus fusiformis and group IIA containing mosquito pathogenic isolates. The phylogenetic tree constructed by taking only variable region was also stable at its nodes with high value of bootstrap. The distinct features specific to each of the species of Lysinibacillus are listed in (Table 4). These taxonomic signatures could be reliably utilized for the screening and identification of mosquito pathogenic strains. Due to the taxonomic value of the unique sequences observed based on variable region in the mosquito-pathogenic strains and phenotypic characters, we propose to designate mosquito- pathogenic strains as a new species Lysinibacillus kychi sp. novo that are hitherto clustered in Group IIA by Krych et al. [9] and Group 1 by Nakamura [16].

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What are Curry Leaves?

Curry leaves, or kadi patta, are the leaves of the curry tree, deductively known as Murraya koenigii Spreng. The plant is local to India and is generally found in tropical and subtropical districts. It is developed in different nations including China, Australia, Nigeria, and Ceylon. 

The stature of the plant ranges from 6 to 15 feet long. The helpful pieces of this plant are its leaves, roots, and bark. The leaves have dependably been looked for after for their one of a kind flavor and handiness in cooking, however there are additionally various medical advantages that make them very engaging. The leaves can be dried or fricasseed, contingent upon the proposed use. The new structure is additionally exceptionally prevalent, both for cooking and home grown medications. 

In Ayurvedic drug, curry leaves are accepted to have a few therapeutic properties: against diabetic, cancer prevention agent, antimicrobial, calming, hostile to cancer-causing, and hepato protective. The roots are utilized for treating body throbs and the bark is utilized for snake nibble alleviation. 

The leaves, with their tremendous home grown properties, are utilized in different nearby foods crosswise over India and Asia as enhancing operators. Curry leaves take after 'neem' or Indian lilac and their name in most Indian dialects means 'sweet neem'.

Curry Leaves Nutrition Facts

The primary supplements found in curry leaves are sugar, fiber, calcium, phosphorous, iron, magnesium, copper, and different minerals.  They additionally contain different nutrients, for example, B3 (nicotinic corrosive), nutrient C, nutrient A, nutrient B, nutrient E, cancer prevention agents, plant sterols, amino acids, glycosides, and flavonoids. 

They additionally have a follow, non-huge measure of fat (0.1 g per 100 g). 

Another concoction constituent present in curry leaves is carbazole alkaloids. An examination concentrate distributed in the Journal of Agricultural and Food Chemistry demonstrated that alkaloids found in the leaves have cancer prevention agent properties. 

Carbazole alkaloids incorporate mahanimbine, murrayanol, mahanineoenimbine, O-methylmurrayamine An, O-methylmahanine, isomahanine, bismahanine and bispyrayafoline. A further report directed at the Department of Horticulture at Michigan State University recommended that these synthetic concoctions have insecticidal and antimicrobial properties too, explicitly mosquitocidal properties.

Benefits of Curry Leaves

A great many people imagine that curry leaves are included for flavor and discard the departs while eating their soup or curry. Be that as it may, they are undeniably more significant than many acknowledge, and offer various medical advantages with no reactions.

  Weight Loss

Curry leaves may help with weight reduction, on account of the nearness of the carbazole alkaloids referenced above, which may forestall weight increase and lower LDL cholesterol. You can include dried or new curry leaves to your nourishment or you can legitimately chomp on the dried curry leaves.

Alleviate Diarrheal

Research demonstrates that the carbazole alkaloids present in curry leaves have antidiarrheal properties. Analyses on guinea pigs demonstrated that carbazole separates from curry leaves altogether controlled castor oil-initiated looseness of the bowels. Expend them by crushing one pack of curry leaves, and eat the glue or the juice of the leaves.

  Treat Indigestion

In Ayurveda, it is thought that the use of curry leaves can be used as a cure for gastrointestinal issues as they are considered to possess mild laxative properties.  You can make juice out of a bunch of curry leaves, add lime juice, and consume the mixture to alleviate indigestion. A paste made from the leaves can also be added to buttermilk and taken every morning on an empty stomach to serve the same function.

  Prevent Nausea & Morning Sickness

It is thought in numerous societies that curry leaves help in anticipating queasiness, regurgitating, and morning affliction. They additionally help give alleviation from morning disorder and queasiness to ladies in their first trimester of pregnancy.

  Fight Infection

Research on curry leaves has uncovered that they are powerful in battling bacterial and parasitic diseases because of the nearness of carbazole alkaloids. This compound is known to have antibacterial, calming, and hostile to disease properties. The leaf separates from the plant have been similar to famous standard anti-infection drugs.

  Anti-diabetic Properties

Maybe one of the greatest medical advantages of curry leaves is their utilization in diabetes control. 

Research referred to in an International Journal of Pharmaceutical Sciences demonstrated that the counter hyperglycemic properties of the leaves were useful in controlling blood glucose level in diabetic rodents.

Good for Eyesight

Curry leaves contain high measures of nutrient A, which is valuable for visual perception, as indicated by specialists. Nutrient A contains carotenoids which ensure the cornea and the eye surface. Lack of nutrient A may cause night visual impairment, cloud developments before the eye, and even loss of vision now and again.

  Fight Oxidative Stress

An exploration concentrate distributed in the diary Food Chemistry has shown that curry leaves are a decent wellspring of cancer prevention agents. [6] The nearness of different nutrients like nutrient A, B, C, and E help in lessening oxidative pressure and free radical rummaging movement.

  Heal Wounds

Curry leaves are likewise useful in healthy skin. The juice or glue of the leaves can be connected to consumes, cuts, wounds, skin disturbances, and creepy crawly chomps for a practical recuperation and clean mending.

  Fight Cancer

The concoction constituents found in curry leaves, for example, phenol's are useful in battling malignant growths, for example, leukemia, prostate disease, and colorectal tumors. One research study indicated proof of colon malignant growth battling properties in the carbazole alkaloids extricates from curry leaves.

  Lower Cholesterol Levels

These leaves have been appeared to effectually affect decreasing LDL or terrible cholesterol levels. Studies have demonstrated that they can possibly lessen LDL cholesterol levels, however further research is expected to affirm the outcomes.

  Hair Care

Curry leaves are accepted to help in reinforcing hair roots. Dry curry leaf powder blended with oil can be connected to your hair. The glue from curry leaves can likewise be connected in instances of silver hair to moderate the turning gray procedure. Doing these all the time can improve hair development too.

  Protect the Liver

 Your liver assumes a noteworthy job in the stomach related procedure. Curry leaves can help offer an increase in security from any assault by free radicals, just as from viral and bacterial assaults that can result in disease. Research on curry leaves has demonstrated that the tannins and carbazole alkaloids present in the leaves displayed great properties.

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Bion Tech Inc. Taiwan receives “Futuristic Inventive Technology Award” for development of innovative biological mosquitocide | AgroNews

"This exhibition focused on the research achievements of innovative technologies from not just agriculture industry, but from all fields and industries nation-wide, including projects such as the application of artificial intelligence, storage of green energy, medicinal biotechnology, nano technology, etc.  The 109 chosen participants were selected nationally during a 3-month evaluation by top…

[medtalksblog]

Reproductive Function Impairment and Tridax Procumbens

Reproductive function issues in men, a prevalent medical condition, can lead to problems like infertility, reduced libido, and erectile dysfunction, impacting sexual health. Factors like hormonal imbalances, physical abnormalities, and lifestyle choices (like smoking and excessive alcohol) contribute to these issues. Thankfully, numerous treatments exist to help men manage and enhance their reproductive function.

Studies by researchers have affirmed that Tridax procumbens leaf extract has the potential to alleviate reproductive function impairments. The extract improves sperm quality, reduces oxidative stress, balances hormones, restores healthy glucose levels, and enhances sexual performance when orally administered.

Tridax procumbens, a wild plant in the Asteraceae family, is prevalent in tropical areas, especially in India. Traditional Ayurvedic medicine has utilized its extracts from leaves, stem, flower, and roots rich in various compounds. Experimental studies show its antioxidant, antibacterial, anti-inflammatory, antimicrobial, and mosquitocidal activities.

The aqueous leaf extract of Tridax procumbens gains attention for potential benefits on reproductive health and sexual performance. A healthy erection requires proper blood circulation, suitable hormone levels, a functional nervous system, and a healthy psychological outlook. Tridax Procumbens positively impacts the body, inducing vasodilation, improving blood circulation, managing erectile dysfunction, boosting sperm production, and increasing serum testosterone concentration.

Compared to Viagra, Tridax Procumbens demonstrated similar effects on male fertility in an experimental study. These findings support its traditional use for sexual well-being. While promising, consulting a doctor before taking any supplement is always advisable, following label instructions.

[mosquitopunks-blog]

Don’t Let Mosquitoes Ruin Your Fun Outing

A fun camping trip is an excellent time for everyone to bond and has great fun. Bugs and mosquitoes can ruin this great adventure. Therefore, after years of scientific research Mosquito Punks have found the way. Our range of aromatic natural mosquito repellent sticks kills and prevents these small insects and bugs from hovering anywhere around you. Some of the features of our incense sticks include: 

 Components Composed From Nature

Our incense sticks are constituted from 100% herbal and natural ingredients. Some of the things we use like Typhaceae fiber, Makko powder, Bamboo stick, Pine resin, Tragacanth for creating each stick are imported from around the world. These superior quality materials ensure that you stay absolutely safe and healthy no matter where you are.

 Soothing Aroma

Our Mosquito repellent lavender incense sticks come in a lot of fragrances that not only repels bugs and mosquitos but create a soothing atmosphere. All our incense sticks can uplift your mood and make you feel relaxed to the core. Apart from the natural upliftment in spirit, these incense sticks efficiently do their job of keeping mosquitoes and bugs at bay.

Long-lasting Effect 

The natural ingredients that constitute our incense sticks make them burn for a relatively long time. This prevents bugs from flying anywhere around you and kills them from the very place they come from. One stick burns for a noteworthy 1 – 1 ½ hour, with its effect lasting for a substantial 20 hours ensuring that you stay protected for a long time. 

DEET Free Formula

DEET is a common ingredient found in most the insect repellents around the world. But DEET has also been found to have an adverse impact on human health in research conducted by independent agencies. Therefore, we make sure that each of our natural mosquito repellent sticks is entirely DEET free so you and your younger ones can stay safe in and around our sticks. 

MosquitoPunks, LLC is a leading supplier of mosquito repellent products. All the products we manufacture are environment-friendly for home and outdoor use. We provide total protection from mosquitoes without the use of pesticides or any other harmful chemicals. Our products make sure that our consumers can enjoy their time indoors or outdoors without any worry of getting infected with any harmful viruses from bugs and insects. Our products are also sold online at Walmart, and Amazon websites.

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CHARACTERIZATION, INSECTICIDAL AND MOSQUITO REPELLENCY PROPERTIES OF ESSENTIAL OILS FROM HYPTIS SUAVEOLENS LEAVES

CHARACTERIZATION, INSECTICIDAL AND MOSQUITO REPELLENCY PROPERTIES OF ESSENTIAL OILS FROM HYPTIS SUAVEOLENS LEAVES

CHARACTERIZATION, INSECTICIDAL AND MOSQUITO REPELLENCY PROPERTIES OF ESSENTIAL OILS FROM HYPTIS SUAVEOLENS LEAVES

  ABSTRACT

Essential oils were extracted from the leaves of Hyptis suaveolens by hydro-distillation, Mosquitocidal effect was done through an experiment devised for the purpose of this research. The effect of the essential oils against mosquito larvae (larvicidal), mosquito repellency…