Partial Sequence Analysis Research Articles

Comprehensive depiction of novel heavy metal tolerant and EPS producing bioluminescent Vibrio alginolyticus PBR1 and V. rotiferianus PBL1 confined from marine organisms

The current study depicts the isolation of luminescent bacteria from fish and squid samples that were collected from Veraval fish harbour. From Indian mackerel, total 14 and from squid, total 23 bioluminescent bacteria were isolated using luminescence agar medium. Two bioluminescent bacteria with highest relative luminescence intensity PBR1 and PBL1 were selected. These two isolates were subjected to detailed biochemical characterization and were tested positive for 5 out of 13 biochemical tests. Furthermore, both PBR1 and PBL1 were able to ferment cellobiose, dextrose, fructose, galactose, maltose, mannose, sucrose and trehalose with acid production. Based on 16S rRNA partial gene sequence analysis, PBR1 was identified as Vibrio alginolyticus and PBL1 as V. rotiferianus. Antibiotic susceptibility test using paper-disc method showed that PBR1 and PBL1 were sensitive to chloramphenicol, ciprofloxacin, co-trimoxazole, gatifloxacin, levofloxacin, linezolid ad roxithromycin out of 18 antibiotics tested. Moreover, both strains were evaluated for their exopolysachharide (EPS) producing ability where PBR1 and PBL1 were able to yield 1.34 g% (w/v) and 2.45 g% (w/v) EPS respectively from 5 g% (v/v) sucrose concentration. Heavy metal toxicity assessment was carried out using agar well diffusion method with eight heavy metals and both the strains were sensitive to As(III), Cd(II), Ce(II), Cr(III), Cu(II), Hg(II) and while they showed resistance to Pb(II) and Sr(II). Based on these results, a study was conducted to demonstrate bio-removal of Pb and Sr by EPS of PBR1 and PBL1. Fourier transform infrared (FTIR) spectra revealed the functional groups of EPS involved in interaction with the heavy metals. Owing to the sensitivity for the remaining heavy metals, these bioluminescent bacteria can be used further for the development of luminescence-based biosensor.

Entomological Surveillance for Zika and Dengue Virus in Aedes Mosquitoes: Implications for Vector Control in Thailand.

Entomological surveillance for arthropod-borne viruses is vital for monitoring vector-borne diseases and informing vector control programs. In this study, we conducted entomological surveillance in Zika virus endemic areas. In Thailand, it is standard protocol to perform mosquito control within 24 h of a reported dengue case. Aedes females were collected within 72 h of case reports from villages with recent Zika–human cases in Kamphaeng Phet Province, Thailand in 2017 and 2018. Mosquitoes were bisected into head-thorax and abdomen and then screened for Zika (ZIKV) and dengue (DENV) viruses using real-time RT-PCR. ZIKV RNA was detected in three samples from two female Ae. aegypti (1.4%). A partial envelope sequence analysis revealed that the ZIKV sequences were the Asian lineage identical to sequences from ZIKV-infected cases reported in Thailand during 2016 and 2017. Dengue virus-1 (DENV-1) and dengue virus-4 (DENV-4) were found in four Ae. aegypti females (2.8%), and partial capsid sequences were nearly identical with DENV-1 and DENV-4 from Thai human cases reported in 2017. Findings in the current study demonstrate the importance of entomological surveillance programs to public health mosquito-borne disease prevention measures and control.

Hydrocarbon-Degrading Bacterial Strain Pseudomonas mendocina Newly Isolated from Marine Sediments and Seawater of Oran Harbor (Algerian Coast)

Contamination by petroleum hydrocarbons causes serious dangers to human health and the environment, whether by accidental or chronic contamination. Due to the large flow of ships, the commercial harbor of Oran is subject to pollution particularly by polycyclic aromatic hydrocarbons. For that, bioremediation by indigenous microorganisms is the most important method to eliminate or decrease this contamination. In the present paper, hydrocarbon-degrading bacterium strain SP57N has been studied, newly isolated from contaminated marine sediments and sea water from the harbor of Oran (Northwestern-Algeria), using of Bushnell-Hass salt medium (BHSM). The strain SP57N was Gram-negative, oxidase negative, catalase negative, motile, Rod-shaped bacteria, identified molecularly as Pseudomonas mendocina based on partial 16S rDNA gene sequence analysis, using the BLAST program on National Centre for Biotechnology Information (NCBI) and the EzBioCloud 16S rDNA databases. This isolate could growth on high concentrations of crude oil (up to 10 %, v/v). The effects of some culture conditions such as temperature, NaCl concentration and pH on growth rate of strain SP57N on crude oil as the sole carbon and energy source were studied. In addition, growth kinetic of this isolate on crude oil during 20 days of culture at 140 rpm, under optimal culture conditions was considered. The results showed maximum growth rate at temperature 25°C, 3% (w/v) of NaCl concentration and pH 7. Results of growth kinetic on crude oil as sole carbon and energy source showed that the stationary phase was attained at day 12. Thus, Pseudomonas mendocina SP57N had effectively hydrocarbon-degrading potential, and could be used as an efficacy degrader to initiate a biological eco-friendly method for the bioremediation of the hydrocarbon pollution on the port of Oran, and marine environment.

Waterborne Outbreak of Acute Gastroenteritis Caused by Recombinant Norovirus GII.P7–GII.6 in Khabarovsk in 2019

Background: Noroviruses are common causative agents of acute gastroenteritis worldwide. Person-to-person transmission is the dominant transmission route for norovirus infection but contaminated water also often leads to outbreaks. Objectives: Our purpose was to do epidemiologic and molecular genetic analyses of waterborne norovirus infection outbreak among children in Khabarovsk in 2019. Materials and methods: Clinical and water samples were screened for the presence of norovirus RNA using real-time RT-PCR detection kit. The norovirus nucleotide sequences were determined by Sanger sequencing. The obtained sequences were subjected to a phylogenetic analysis. Results: In July 2019, 34 children developed acute gastroenteritis in Khabarovsk. The epidemiologic investigation showed that on the eve of the disease onset all patients played and bathed in a pedestrian fountain complex. A molecular genetic analysis of 18 biological samples from children with acute gastroenteritis and a water sample from the fountain revealed a recombinant norovirus GII.P7-GII.6. We established a 100.0% identity of all obtained nucleotide sequences to each other. A phylogenetic analysis of ORF2 partial sequences showed that the capsid protein of the Khabarovsk GII.P7-GII.6 strains belonged to the variant GII.6a. Conclusions: Contaminated water in the pedestrian interactive fountain complex was the most likely cause of the norovirus infection outbreak among children in Khabarovsk in 2019 associated with the lack of proper maintenance and regular disinfection measures.

Diversity and pathogenicity of Botryosphaeriaceae species associated with black root rot and stem cutting dry rot in Manihot esculenta in Brazil

Cassava is a starch-rich, high-calorie tuberous root used as human and animal food, as well as an input material in various industries. Black root rot (BRR) and stem cutting dry rot (SCDR) are cassava diseases that compromise product quality and can greatly reduce crop yields. Consequently, the aim of this study was to identify Botryosphaeriaceae species associated with both diseases in areas of the northeast of Brazil and in areas of the Minas Gerais State in the southeast of the country and infer a correlation between them. Fungal species were identified by morphology combined with phylogenetic analyses of partial sequences of translation elongation factor 1 alpha (tef1-α), internal transcribed spacer (ITS) and RNA polymerase subunit II (rpb2). Five species were identified as being associated with both BRR and SCDR (Lasiodiplodia euphorbicola, L. hormozganensis, L. parva, L. theobromae, and Neoscytalidium dimidiatum), while five others were associated only with SCDR (L. brasiliense, L. caatinguensis, L. iraniensis, L. laeliocattleyae, and L. pseudotheobromae). All species identified in this survey were able to induce disease in cassava roots and seedlings. In decreasing order, the most frequently isolated species were L. theobromae, N. dimidiatum, and L. euphorbicola. This is the first report of L. brasiliense, L. caatinguensis, L. hormozganensis, L. iraniensis, L. laeliocattleyae, and L. parva associated with cassava diseases worldwide.

Drought Stress Tolerance and Photosynthetic Activity of Alloplasmic Lines T. dicoccum x T. aestivum.

Tetraploid species T. dicoccum Shuebl is a potential source of drought tolerance for cultivated wheat, including common wheat. This paper describes the genotyping of nine stable allolines isolated in the offspring from crossing of T. dicoccum x T. aestivum L. using 21 microsatellite (simple sequence repeats—SSR) markers and two cytoplasmic mitochondrial markers to orf256, rps19-p genes; evaluation of drought tolerance of allolines at different stages of ontogenesis (growth parameters, relative water content, quantum efficiency of Photosystem II, electron transport rate, energy dissipated in Photosystem II); and the study of drought tolerance regulator gene Dreb-1 with allele-specific PCR (AS-MARKER) and partial sequence analysis. Most allolines differ in genomic composition and T. dicoccum introgressions. Four allolines—D-b-05, D-d-05, D-d-05b, and D-41-05—revealed signs of drought tolerance of varying degrees. The more drought tolerant D-41-05 line was also characterized by Dreb-B1 allele introgression from T. dicoccum. A number of non-specific patterns and significant differences in allolines in regulation of physiological parameters in drought conditions is identified. Changes in photosynthetic activity in stress-drought are shown to reflect the level of drought tolerance of the forms studied. The contribution of different combinations of nuclear/cytoplasmic genome and alleles of Dreb-1 gene in allolines to the formation of stress tolerance and photosynthetic activity is discussed.

Genome constellations of rotavirus a isolated from avian species in Brazil, 2008-2015.

Rotaviruses are members of the family Reoviridae and are a common cause of acute diarrhea in many mammalian and avian species. They are non-enveloped icosahedral particles and their genome comprises 11 segments of double-stranded RNA, which encodes six structural proteins (VP1-4, VP6-7) and six nonstructural proteins (NSP1-6). Genotypes are defined based upon the diversity found in these genes and viral characterization plays a central role on epidemiological studies and prevention. Here we investigate the distribution of Brazilian RVAs genotypes in 8 chicken samples collected between 2008 and 2015 from different regions by RT-PCR, partial (Sanger) nucleotide sequencing and phylogenetic analysis from all rotavirus genes. Although the identified genotypes were typical from avian host species, when analyzed together, they form novel genetic constellations: G19-P[31]-I11-R6-C6-M7-A16-N6-T8-E10-H8 and G19-P[31]-I4-R4-C4-M4-A16-N4-T4-E4-H4. This study highlights that avian rotaviruses are widespread among commercial farms in Brazil, and the co-circulation of at least two different genomic constellations indicates that may present a way bigger genetic variability, that can be increased by the possible transmission events from other birds, lack of specific preventive measures, as well as the different viral evolution mechanisms.

Spatial genetic structure of black francolin (Francolinus francolinus asiae) in the North-Western Himalayan region based on mitochondrial control region

Due to specific habitat preferences and behavioural limitations, black francolin is not uniformly distributed across the northwestern Himalayan landscape, rather is confined to certain land mosaic. The habitable zones are further reduced due to several manmade threats as logging and forest fire leading to sparse distribution. Overall 54 samples were used for partial sequence analysis of mitochondrial control region. A well evident divergence pattern was observed as individuals collected from low altitude, terrai region significantly distanced from high altitude sampled individuals. Also, the individuals at lower elevation sites exhibited higher genetic diversity in comparison to the samples collected at higher elevations. This indicates that patchy distribution and low dispersal rate have resulted in fine-scale patterns of genetic diversity among the black francolin population. Further, habitat loss and forest fragmentation could lead to more small and isolated populations that could suffer from reduced genetic diversity and may be higher extinction rates.

Bacteria producing antimicrobials against Clostridium difficile isolated from human stool

Clostridium difficile infection (CDI) is a common cause of morbidity and mortality in hospitalized patients worldwide. The major problem facing current treatment is multiple recurrences, prompting the need for alternative therapies. In this study we isolated bacterial species, from Egyptian individuals’ stool, with antimicrobial activity against clinical isolates of C. difficile and tried to examine the nature of the produced antimicrobials. In vitro antibacterial activity against C. difficile was initially screened in 123 fecal samples cultures using an agar overlay method. The isolates with antimicrobial activity against C. difficile in addition to Clostridium isolates were identified using partial 16S rDNA gene sequencing analysis. The isolates acting against C. difficile belonged to Lactobacillus, Enterococcus and Clostridium genera. The concentrated cell-free supernatants (CFSs) from these bacterial isolates were examined for antimicrobial activity against C. difficile growth by broth dilution method. 10 x concentrated CFSs of five isolates showed inhibition for C. difficile growth which was significantly different (p < 0.001) from control. Lactobacillus agilis T99A and Clostridium butyricum T58A isolates were selected for further evaluation of the produced antimicrobials. The antimicrobial activity of 10x CFSs of the two isolates was stable after enzymatic treatment with proteinase K or heating treatments up to 90 °C or neutralizing pH. The spectrum of activity of the two isolates was evaluated using different gram-positive and gram-negative bacterial species and did not show antimicrobial activity against these species. Our results showed two unconventional bacterial isolates: L. agilis T99A and C. butyricum T58A producing extracellular thermo stable antimicrobial agents against C. difficile clinical isolates.

Sensitivity of Colletotrichum acutatum Isolates from Citrus to Carbendazim, Difenoconazole, Tebuconazole, and Trifloxystrobin.

Postbloom fruit drop (PFD) of citrus is caused by the Colletotrichum acutatum and C. gloeosporioides species complexes. The disease is important when frequent rainfall occurs during the flowering period of citrus trees. In Brazil, until 2012, PFD was mainly controlled by preventive applications of the methyl-benzimidazole carbamate (MBC) carbendazim and demethylation-inhibitor (DMI) fungicides such as difenoconazole. Since then, mixtures containing the DMI tebuconazole and the quinone-outside inhibitor (QoI) trifloxystrobin have been commonly used. Fungicides are often applied preventively, sometimes even when conditions are not conducive for PFD development. Excessive fungicide applications may favor the selection of resistant populations of Colletotrichum spp. In this study, we assessed the fungicide sensitivity of C. acutatum isolates collected during the two distinct periods of PFD management in Brazil: before and after the trifloxystrobin and tebuconazole mixture became widely employed. The sensitivity of 254 C. acutatum isolates to carbendazim and difenoconazole and of 164 isolates to tebuconazole and trifloxystrobin was assessed. Mycelial growth inhibition of these isolates was evaluated for all the fungicides using either serial dilution of fungicide rates or the spiral gradient dilution method. In addition, inhibition of conidial germination was also assessed for trifloxystrobin. Analysis of partial β-tub, cytb, and cyp51b gene sequences did not reveal any mutations related to resistance to MBCs, QoIs, and DMIs, respectively. In mycelial growth assays, mean EC50 values were 0.14, 0.11, and 0.21 μg/ml for difenoconazole, tebuconazole, and trifloxystrobin, respectively. The conidial germination inhibition by trifloxystrobin was similar among the tested isolates, and the mean EC50 value was 0.002 μg/ml. All isolates had similar mean mycelial growth inhibition for carbendazim, regardless of the fungicide concentrations. Therefore, based on similar EC50 values and molecular analyses, no shift in the sensitivity of isolates has been observed to the fungicides commonly used in different citrus-producing areas in Brazil.

Process optimization and characterization of polyhydroxyalkanoate copolymers produced by marine Pichia kudriavzevii VIT-NN02 using banana peels and chicken feather hydrolysate

Abstract The present study was focused on the production of polyhydroxyalkanoates (PHA) by halophilic yeast using biowaste materials. The yeast strain was isolated from marine seaweeds and identified as Pichia kudriavzevii VIT-NN02 based on partial 18SrDNA sequencing and phylogenetic analysis. PHA granules were observed by Sudan Black B and Nile Red staining methods. Banana peels and chicken feathers hydrolysate were screened as potential carbon and nitrogen sources for PHA content in biomass (%) by P. kudriavzevii. Process parameters were optimized using 3-level, six variables, Box-Behnken design showing significant improvement in PHA content in biomass from 40.00 ± 0.1 to 79.68 ± 0.2% under optimized growth conditions viz. banana peel extract (40%), salt (16.6gL-1), incubation time (96 h), pH (10), temperature (37 °C) and chicken feather hydrolysate (0.80%). The statistical significance of the model equation was calculated by F-test and R2 values of the respective responses were found to be 0.9904 and 0.9926, indicating a realistic fit of the model to the experimental data. The actual values of the PHA content in biomass (79.68 ± 0.2%) was in close agreement to the predicted value (79.68 ± 1.5%), confirmed the validity of the model. The biopolymer was identified as poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] copolymer based on Gas Chromatography-Mass Spectrometry (GC-MS), Fourier Transform Infrared (FTIR) spectroscopy and Nuclear Magnetic Resonance spectroscopy (NMR) analysis. The [P(3HB-co-3HV)] copolymer was found to be highly crystalline and thermally stable through X-Ray Diffraction (XRD), Thermogravimetric analysis (TGA) and Differential scanning calorimetric (DSC) analysis. The Scanning Electron Microscopic (SEM) images of [P(3HB-co-3HV)] copolymer showed porous, smooth and spherical surface morphology.

Endosymbiotic pathogen-inhibitory gut bacteria in three Indian Major Carps under polyculture system: A step toward making a probiotics consortium

The gastrointestinal (GI) microbiome in fish plays significant roles in health and disease resistance. This investigation was accomplished to enumerate, characterize and identify the potential probiotic bacteria from three Indian Major Carps (IMCs), viz., rohu (Labeo rohita), catla (Catla catla) and mrigal (Cirrhinus mrigala) using culture dependent methods. Altogether, 105 pathogen-inhibitory bacteria (out of 1216 isolates) were detected from three IMCs by double layer assay. 16S rRNA partial gene sequence analyses and BLAST search in the NCBI GenBank unveiled that 94.29% of the pathogen inhibitory bacteria were bacilli (99 strains) and Bacillus licheniformis by far the most common (28%). The primarily selected 27 pathogen-inhibitory strains (cumulative inhibition score ≥13) produced extracellular enzymes, while 15 of them produced all the six exo-enzymes studied (amylase, protease, lipase, cellulase, phytase and xylanase). Gut stability of the strains became apparent by their ability to grow in fish mucus and tolerance to diluted bile-juice. Finally, 14 strains were noticed as γ-hemolytic and susceptible to the commonly used antibiotics. Further, intra-peritoneal injection with γ-hemolytic strains did not induce any pathological signs or mortalities in fish, and thus were considered as safe. These 14 γ-hemolytic isolates were represented by the genus Bacillus (13) and Stenotrophomonas (1), which might form probiotic consortia for prospective use in carp culture.

Evidence for population expansion of Cotton pink bollworm Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) in India

Pink bollworm, Pectinophora gossypiella (Saunders) infestation on Bt cotton is a major concern to cotton production in India. The genetic diversity and phylogeographic structure of the insect in light of PBW resistance needs to be revisited. The objective of this study was to identify different haplotypes of pink bollworm and their distribution in India. To achieve this we studied the population structure in 44 cotton growing districts of India. The partial mitochondrial COI sequence analyses of 214 pink bollworm populations collected from 44 geographical locations representing 9 cotton growing states of India were analysed. Genetic diversity analysis exhibited presence of 27 haplotypes, among them Pg_H1 and Pg_H2 were the most common and were present in 143 and 32 populations, respectively. Distributions of pairwise differences obtained with partial COI gene data from the overall Indian populations are unimodal, suggesting population expansion in India. Significant neutrality test on the basis of Tajima’ D and Fu’s Fs presented a star-shaped haplotype network together with multiple haplotypes. The unimodal mismatch distribution, rejection of neutrality test with significant negative values supported the theory of demographic expansion in cotton pink bollworm populations in India. Genetic data not only provides us with a perspective of population genetics, but also that the two populations of pink bollworm, those occurring early in the season are genetically close to the late season populations with respect to their partial CO1 region. Resistance to Cry toxins does not seem to have had an impact on this region of the mt DNA in populations of pink bollworm.

Optimization of carbofuran insecticide degradation by Enterobacter sp. using response surface methodology (RSM)

Response surface methodology and Plackett-Burman experiments were applied to optimize the biodegradation of carbofuran by using Enterobacter sp. stain BRC05 isolated from selected agricultural areas in peninsular Malaysia. The significant factors influencing the degradation of carbofuran were assessed using two-level Plackett–Burman Design (PBD) with five variables. Plakett Burman experiment showed that the following four variables were significant for carbofuran degradation including, carbofuran concentration, temperature, pH and nitrogen sources. Significant variables obtained in Plackett-Burman Design were further optimize using Central Composite Design (CCD). The outcome of the design for carbofuran degradation for each runs of the PBD experiment base on the design matrix, showed that the minimum and the maximum carbofuran degradation percentage were found to be 6.7% and 79.77% as presented in runs 4 and 1, respectively. Results obtained using Central Composite Design showed that the relations between the factors affect carbofuran degradation with significant response. The predicted results in CCD indicated that highest carbofuran degradation of 95.40% could be realized with carbofuran concentration of 92.50 mg/L, pH of 6.0, temperature 27.50 °C, nitrogen sources of 0.45 g/L and reaction period of 6 days. The predicted values were in agreement with the actual values with coefficient of determination with R2 0.9719. Partial 16S rRNA sequence analysis showed that the carbofuran degrading isolate was closely related to members of the genera Enterobacter sp. The morphological and biochemical characteristics of the isolate also confirmed the phylogenetic signature. This study would provide an effective approach that could be beneficial for the bioremediation of carbofuran insecticide in polluted environment.

No bejel among Surinamese, Antillean and Dutch syphilis diagnosed patients in Amsterdam between 2006–2018 evidenced by multi-locus sequence typing of Treponema pallidum isolates

Treponema pallidum subspecies pallidum (TPA) and subsp. endemicum (TEN) are the causative agents of syphilis and bejel, respectively. TEN shows similar clinical manifestations and is morphologically and serologically indistinguishable from TPA. Recently, bejel was found outside of its assumed endemic areas. Using molecular typing we aimed to discover bejel and characterize circulating TPA types among syphilis cases with Surinamese, Antillean and Dutch ethnicity in Amsterdam. DNA was extracted from 137 ulcer swabs, which tested positive in the in-house diagnostic PCR targeting the polA gene. Samples were collected between 2006 and 2018 from Surinamese, Antillean and Dutch patients attending the Amsterdam STI clinic. Multilocus sequence typing was performed by partial sequence analysis of the tp0136, tp0548 and tp0705 genes. In addition, the 23S rRNA loci were analyzed for A2058G and A2059G macrolide resistance mutations. We found 17 distinct allelic profiles in 103/137 (75%) fully typed samples, which were all TPA and none TEN. Of the strains, 82.5% were SS14-like and 17.5% Nichols-like. The prevalence of Nichols-like strains found in this study is relatively high compared to nearby countries. The most prevalent types were 1.3.1 (42%) and 1.1.1 (19%), in concordance with similar TPA typing studies. The majority of the TPA types found were unique per country. New allelic types (7) and profiles (10) were found. The successfully sequenced 23S rRNA loci from 123/137 (90%) samples showed the presence of 79% A2058G and 2% A2059G mutations. No TEN was found in the samples from different ethnicities residing in Amsterdam, the Netherlands, so no misdiagnoses occurred. Bejel has thus not (yet) spread as a sexually transmitted disease in the Netherlands. The strain diversity found in this study reflects the local male STI clinic population which is a diverse, mixed group.

New evidence of bovine leukemia virus circulating in Myanmar cattle through epidemiological and molecular characterization.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, which is the most common neoplastic disease of cattle. BLV infects cattle worldwide and causes serious problems for the cattle industry. In this study, we examined the prevalence of BLV infection and the distribution of BLV genotypes in cattle in the northern, central, and southern parts of Myanmar. The prevalence of BLV infection among Myanmar cattle (37.04%) in this study was markedly higher than the prevalence (9.1%) observed in our earlier study in which BLV was detected from the limited number of cattle only from a small area of Myanmar. Phylogenetic analysis of partial env-gp51 sequence of the isolated BLV strains revealed that there are at least three BLV genotypes (genotype-1, genotype-6, and genotype-10) in Myanmar, which have also been detected in the neighboring countries. We performed this study to estimate the BLV proviral load, which is a major diagnosis index for determining the virus transmission risk. The cattle of the three test regions with warm, wet, and humid climatic conditions (upper Sagaing, Yangon, and Kayin) exhibited a high mean proviral load, while cattle of three other regions with low annual rainfall and very high temperature (Mandalay, Magway, and upper Bago) exhibited a low mean proviral load. Further, the level of proviral load and the prevalence of BLV infection in Myanmar native cattle (N = 235) were lower than that in the hybrid cattle (Holstein Friesian × Myanmar native) (N = 62). We also observed that the cattle with high risk for BLV transmission, which have high proviral load, may enhance the BLV infection rate. Hence, to control BLV transmission, it is necessary to eliminate these cattle with high-risk for BLV transmission and to diagnose BLV provirus in cattle in the remaining regions/states of Myanmar sharing a boundary with neighboring countries.

A New Species of Triumphalisnema (Nematoda: Coronostomatoidea) Parasite of Proculejus hirtus (Coleoptera: Passalidae) from Mexico, and the Position of the Family in the Oxyuridomorpha

A new species of the nematode Triumphalisnema Kloss, 1962 (Oxyuridomorpha), is described from the wood beetle Proculejus hirtus Truqui from the mountain mesophilic forest in Hidalgo State, Mexico. Triumphalisnema zuuei n. sp. is distinguished from the other 4 congeners species by the presence of an expanded cervical ring, well-developed lateral alae, an obtuse cauda with a short and bifurcated caudal appendage, series of cuticular folds at ventral and dorsal body surface from excretory pore level to anal region, and ellipsoidal eggs ornamented with numerous small mushroom-like structures over the surface. The phylogenetic position of the new species is inferred based on a Maximum Likelihood and Bayesian Inference analysis of partial sequences of 18S SSU rRNA. The phylogenetic analysis showed that Triumphalisnema zuuei n. sp., the only representative of the Traklosiidae in our study, is closely related to Coynema poeyi and species of Longior, Hystrignathus, and Lepidonema, all of them members of Hystrignathidae. These relationships are supported by high support values. The present study increases to 5 the number of species assigned to Triumphalisnema, all of them parasites of Passalidae. Additionally, a taxonomic key to the species of the genus is provided.

SXT int harboring bacteria as effective indicators to determine high-risk reservoirs of multiple antibiotic resistance in different aquatic environments of western coast of Gujarat, India

Unregulated use of antibiotics in the developing countries has led to the development of Multiple Antibiotic Resistant (MAR) strains. Resistance transferring elements like SXT play crucial role in dissemination of the resistance genes. Persistence of antibiotics and their resistance elements in an aquatic ecosystem depends on the physicochemical properties, and thus may depend on the type of whole water body in general. The study focuses on isolation of bacteria possessing SXT integrase from diverse aquatic bodies like polluted sources, freshwater ponds, groundwater and seawaters of western Gujarat coast, and studying distribution of multidrug resistance in them. Initially, 246 Co-Trimoxazole resistant microbes were isolated from various environmental samples on the basis of morphological distinction. In the PCR analysis, 45 of them were found to be positive for the SXT integrase gene and subsequent analysis confirmed the presence of SXT or SXT-variants. Partial 16S rRNA gene sequencing and diversity analysis on the basis of Shannon’s diversity indices revealed bacterial class-wise diversity trend to be pond water > seawaters > ground water > polluted sources. Antibiogram of these organisms revealed the order of antibiotic resistance indices to be polluted sources > groundwater sources > freshwater ponds > seawater sources. However, the trend of percentage resistance towards antibiotic classes separately in the studied aquatic environments was groundwater sources > polluted sources > freshwater ponds > seawater. Further, the trend for therapeutic efficiency of antibiotic classes was similar in all waters to that of total isolates i.e. Aminoglycosides > Fluoroquinolones > Amphenicols > Cephalosporins > Imidazolidinedione derivatives > Glycopeptide > Quinolones > Macrolides > Penicillins > Sulfa drugs. Positive correlation was observed between prevalence of SXT int positive bacteria and multiple antibiotic resistance incidences; but negative correlation was observed between SXT int positive bacteria and class wise bacterial diversity in four designated type of aquatic environment. Thus, screening for occurrence frequency of SXT integrase gene in the bacterial isolates, may prove as an effective parameter to estimate the multiple antibiotics’ resistance indices in any of the four studies aquatic ecosystems, and thereby determine the antibiotic pollution resulting in the corresponding pressure. The overall quantitative values were logically higher in polluted sources in terms of total resistance indices, and freshwater ponds in terms of diversity. But the groundwater sites may be regarded as the prominent high-risk zones of contamination with multiple antibiotic and/or their resistance traits qualitatively.

Stepwise-Selected Bacillus amyloliquefaciens and B. subtilis Strains from Composted Aromatic Plant Waste Able to Control Soil-Borne Diseases

In the present study, 133 bacterial isolates from 11 composted aromatic plant wastes were selected for their ability to inhibit the mycelial growth of the soil-borne phytopathogenic fungi Sclerotinia minor and Rhizoctonia solani. Successively, a subset of 35 from them were further characterized for their ability to control, in vivo, rocket damping-off caused by the two fungi. Moreover, the isolates were characterized for morphology of the colonies, Gram reaction, siderophore production, P-solubilization and for the presence of antimicrobial lipopeptide genes in the genome. The screening for the in vitro antagonisms showed a mycelial growth reduction ranging between 31.7% and 56.1% for R. solani and 34.4% and 59.4% for S. minor. All the isolates were not able to produce siderophores and some of them were able to solubilize P. The isolates contained two or more of the five lipoproteins coding genes investigated in this study. The most promising isolates were identified at species level by 16S-rRNA partial gene sequence analysis and were grouped in two main clusters related to Bacillus subtilis and Bacillus amyloliquefaciens reference strains. Results indicated that Bacillus isolates from compost are good candidates for application in the biocontrol of cultivated plants.

Sequence analysis of nucleoprotein gene reveals the co-circulation of lineages and sublineages of rabies virus in herbivorous in Rio Grande do Sul state, Brazil.

An unprecedented outbreak of rabies occurred in Rio Grande do Sul state (RS) from 2012 onward, resulting in thousands of bovine deaths, important economic losses, and posing risk to human health. This article describes a genetic analysis of 145 rabies viruses (RABV) recovered from herbivorous from RS between 2012 and 2017, based on partial sequence analysis of the nucleoprotein (N) gene. High nucleotide (nt) identity (95.5 to 100%) and amino acid (aa) similarity (96.7 to 100%) were observed among the analyzed sequences. These sequences displayed a high sequence nt identity/aa similarity with bovine RABV sequences (96.4-97.9%; 98.1-100%, respectively) and vampire bat RABV sequences (96.3-97.5%; 97.8-99.5%). Phylogenetic analyzes based on the N sequence allowed for the segregation of viruses into two distinct clusters. Cluster 1 comprised RABV sequences covering the whole studied period, whereas cluster 2 grouped a lower number of viruses from 2013, 2014, 2015, to 2017. In some cases, viruses obtained from the same region within a short period of time grouped to distinct clusters or sub-clusters, indicating the co-circulation of distinct virus lineages in these outbreaks. The segregation into sub-clusters was also observed for viral sequences obtained from the same region at different times, indicating the involvement of distinct viruses. In summary, partial sequence analyses revealed a high conservation of N protein and the circulation of two lineages and different sublineages of RABV in the region. In addition, our results confirm the suitability of N gene to study the genetic relationships among RABV isolates.