Background: Escherichia coli and Klebsiella pneumoniae are two important commensals with pathogenic potentiality in animals and birds, causing intestinal and extra-intestinal infections. These are somewhat zoonotic too via different routes. So, the present study was undertaken to investigate the prevalence of these two bacteria from cloaca and environment of ducks from different districts of West Bengal, followed by detailed characterization and antibiogram. Methods: Around 197 duck and environmental samples were collected from different districts of West Bengal. E. coli and K. pneumoniae were isolated from collected samples as per standard methods followed by serotyping of the E. coli isolates. The detection of ESBL/ACBL producing isolates was done both phenotypically and genotypically by detecting the presence of antibiotic-resistant genes. Virulence property and biofilm-producing ability of these isolates were also studied. Antibiogram of the ESBL and biofilm positive isolates were done using common 12 antibiotics by disc diffusion method. Result: A total of 134(68.02%) Escherichia coli and 33(16.75%) Klebsiella pneumoniae isolates were recovered and confirmed by PCR. The prevalence of E. coli and K. pneumoniae in cloacal swabs were less in comparison to the environmental samples (72.09% and 20.93%). Approx. 79.85% E. coli and 72.73% K. pneumoniae isolates were found to be the ESBL/ACBL producers in vitro. E. coli and K. pneumoniae isolates possessed all major antibiotic-resistant genes with the blaAmpC (64.18% and 48.48%) being the most prevalent one. Twenty three (17.16%) E. coli isolates were found positive for the virulence genes, of which stx1 (39.13%) was the most common one as detected by multiplex PCR. No virulence gene was found in K. pneumoniae isolates. Ninety one (67.91%) E. coli and 26(78.79%) K. pneumoniae isolates possessed the biofilm-producing genes. All the ESBL and biofilms producing E. coli and K. pneumoniae isolates showed 100% - 95.24% resistance to ampicillin. A high percentage of resistances were also seen against cefotaxime, ticarcillin/ clavulanic acid, ertapenem, ceftazidime, norfloxacin and ciprofloxacin, which may be due to several reasons. Maximum overall sensitivity was detected against chloramphenicol, co-trimoxazole and gentamicin.

Escherichia coli is quite a common and sometimes pathogenic bacterium that can cause acute to severe infection in poultry birds affecting their performance. E. coli can also cause a few extra-intestinal infections too in poultry birds. Nowadays antimicrobial resistance is quite a menace which is reported from several gram-negative bacteria including E. coli. These traits in E. coli are governed by a few & beta;-lactamases producing genes which may also be associated with plasmids in the pathogen. This study aimed to detect the presence of extended-spectrum (ESBL) and AmpC (ACBL) beta-lactamases producing E. Coli from fecal samples of diseased and apparently healthy poultry birds from different districts of West Bengal, India. A total of 177 (62.32%) E. coli isolates were detected and confirmed from the poultry fecal samples/ cloacal swabs (n=284) collected in this study. All isolates were typical in morphology. One hundred thirty-two (74.58%) bacterial isolates were positive for phenotypical beta-lactamases production, of which 83 (46.89%) were positive for phenotypical ACBL production. A total of 41 and 17 isolates were found to be positive for PCR detection of the blaCTXM and blaSHV genes respectively. No blaTEM gene was detected. Again 95 (53.67%) isolates possessed the blaAmpC gene as detected in this study. Antibiogram of the ESBLproducing E. coli isolates (49) revealed a significantly high level of resistance against most of the commonly used antimicrobials like ceftriaxone, ampicillin (both 100%), cefotaxime (97.96%), ceftazidime (91.84%) amoxicillin/clavulanic acid (85.71%), azithromycin, tetracycline, norfloxacin (@70-75%). These isolates were found to be sensitive (@ 96-81%) against a few antimicrobials, viz. amikacin, imipenem, gentamicin, and ampicillin/sulbactam which is a major point of concern.

The generation of antimicrobial-resistant bacteria largely depends on the use of antimicrobials not only in humans but also in pet animals and livestock. The present study was conducted to detect the occurrence of beta-lactamase and biofilm-producing-E.coli in healthy pet and backyard livestock. The study also intended on molecular docking experiments to confirm the nature of the catalytic mechanism in 8-lactamase enzymes, encoded by the various blaCTX-M genotypes and phylogenetic analysis to reveal clonal relationship of the animal origin E. coli isolates with human clinical strains. The rectal swabs were collected from healthy dogs (n = 254), cats (n = 108), sheep (n = 119) and goats (n = 143) in India. In total 247 (76.47%) E. coli strains were identified as ESBL producers. The possession of ESBL-producers was significantly more (p < 0.05) in pets than in the backyard livestock. Most of the strains possessed blaCTX-M-15 like clones. E. coli strains possessing blaCTX-M-15.2, blaCTX-M-157, blaCTX-M-181 and blaCTX-M-218 like clones, isolated from pets were not reported earlier. The study detected 56.65% of E. coli strains as moderate or strong biofilm producers possessing biofilm-associated genes (csgA, rcsA, rpoS, sdiA). ESBL-producing E. coli showed phenotypical resistance to tetracycline (93.1%), azi-thromycin (89.8%), ampicillin (84.2%), cefotaxime (80.9%), doxycycline (82.5%), co-trimoxazole (80.9%), ampicillin/cloxacillin (76.9%). The CTX-M variants obtained in this study were modelled by the SWISS-MODEL and verified. Ligand having minimum binding energy, show the highest affinity of 8-lactamases for cefotaxime and cefpodoxime. The Gibbs free energy release for all 14 different complex ranges between-6.9 (CTX-M-15.2+cefpodoxime) to-5.3 (CTX-M-218+cefpodoxime) Kcal/mol. Phylogenetic analysis of the animal origin ESBL-E. coli strains revealed a partial clonal relationship with the clinical isolates of local human patients. The present study described the significant presence of biofilm and 8-lactamase producing, multi-drug resistant E. coli in pet animals having public health importance.

Background To study the immunomodulatory and protective role of dietary administered purified β-glucan obtained from edible mushroom ( Pleurotus florida ) in commercial broiler chicken, experimentally challenged with virulent Newcastle disease virus (NDV) on 7th day post treatment. Mushroom glucan (MG) at 15 mg/kg feed (group A) and MG at 30 mg/kg feed (group B) was administered to broiler birds for 20 days keeping control birds (group C) with a normal diet throughout. After 7 days post treatment, three groups of birds ( n = 4, in each case) were challenged with virulent NDV. The immunological parameters were assessed to observe the protective efficacy of MG. Results When compared to the treatment regime, it was observed that in all the cases, group B birds showed higher immune-cellular and humoral responses in terms of enhanced immune-effector activities of blood leucocytes and intestinal intra-epithelial leucocytes and antibody production besides protection against NDV challenge than the others. After NDV challenge, 100% mortality was observed in control birds within 4 days, whereas in treated birds 50% and 75% protection of challenged birds was observed in group A and group B birds, respectively. The superoxide anion production by blood leucocytes of group A (0.641 ± 0.01) and group B (0.721 ± 0.01) birds were significantly higher than the control birds (0.283 ± 0.04) when assessed on 4th day post challenge. Group A (27.33 ± 1.20 μl and 25.33 ± 2.02 μl) and group B (33.66 ± 0.33 μl and 32.66 ± 0.33 μl) birds showed higher in vitro nitrite production by peripheral blood mononuclear cells (PBMC) and intestinal intra-epithelial leucocytes (iIEL), respectively, than the control (14.00 ± 0.57 μl and11 ± 0.57 μl) after challenge with virulent ND virus. In vitro lymphoproliferation (expressed as stimulation index) was significantly high in PBMC and iIEL of group A (0.371 ± 0.02 and 0.295 ± 0.02) and group B (0.428 ± 0.01 and 0.314 ± 0.01), respectively, than control (0.203 ± 0.01 and 0.135 ± 0.01) on 4th day of NDV challenge. The phagocytic activity of iIEL of the treated group birds showed higher values (24% and 32%) than the control group (14%). The haemagglutination inhibition (HI) titre was also observed higher in treated groups (group A, average HI titre 256, and group B, average HI titre 512) than control (HI titre, 32). Both groups (A&B) of birds were produced in vitro IFN-γ by PBMC and iIEL. Conclusion It is advisable to use 30 mg MG/kg feed in broiler birds to provide immunostimulation and for better output in terms of disease protection at least against ND virus.

COVID-19 is nowadays a highly infectious disease affecting human civilization throughout the Globe including India. A novel Coronavirus (nCoV) which was first reported from Wuhan, People's Republic of China, has now emerged as a public health emergency of international concern. The pandemic is caused by the Novel virus of the Coronaviridae family known as SARS-CoV-2. The outbreak was first reported in December 2019, and at present has spread in almost all over the world with more than 23 million cases causing more than 8 lakh 9 thousand deaths up to August 2020. At present India is having more than 3 million confirmed cases (3rd highest in the infection list) of novel coronavirus disease 2019 (COVID-19), with 56000+ deaths, resulting in an overall case fatality rate of 2.04 percent. By August 2020, 215 countries around the world are under the threat of the pandemic with a death rate of 5% approx. Severe acute respiratory syndrome 2 is generally transmitted via close contact with active and non-symptomatic patients and also via air droplets or aerosol routes. The virus generally infects the respiratory system and later on goes on affecting all vital organs in humans. The development of acute respiratory tract infection with fever, cough, diarrhoea, dyspnoea, headache, and breathing trouble are a few typical signs of this infection. In this background, this review will focus on virology, the nature of the infection and epidemiology, pathogenicity, symptomology, diagnostic techniques, possible remedies, and scope to overcome this threat over the mankind with special reference to the Indian perspective. This review will also focus on the key ongoing efforts to prevent the COVID-19 outbreak and identifies the needs to be incorporated in health management systems, and community response mechanisms towards improving global as well as Indian health security.

Escherichia coli are important commensal with pathogenic potentiality in bovine neonates and may cause intestinal and extra-intestinal infections.Different enteric pathogens such as E. coli may remain associated with bovine diarrhoea which is a multifactorial menace.Possession of antimicrobial resistance genes by these commensal or pathogens associated with diarrhea makes the issue more serious.The study was aimed to detect the occurrence and characterize extended-spectrum (ESBL) and AmpC (ACBL) beta-lactamases producing pathogenic E. coli (STEC) in diarrhoeal faecal samples collected from bovines.All the E. coli strains (13/21, 61.9%) isolated from bovine diarrhoea cases (n=21) were found to be ESBL producers.In total, 11 (84.6%)isolates were positive in cefoxitincloxacillin double-disc synergy (CC-DDS) test for AmpC production and the isolates were also confirmed by PCR for bla AmpC .Any of the E. coli isolates were not carrying the class I integrons but 7 (53.8%) of those isolates were found to possess theSTEC gene, eaeA (responsible for causing diarrhoea) as revealed in PCR assays.All the ESBL producing E. coli isolates showed a high level of resistance to amoxicillin/clavulanic acid, ceftriaxone, ceftazidime, ampicillin/cloxacillin, cefotaxime, cefepime, cefoperazone (100%), cefoxitin (84.6%) and tetracycline (61.5%) whereas Ertapenem (92.3%), and enrofloxacin (84.6%) were intermediately sensitive against these pathogens.All the isolates were susceptible to amikacin, levofloxacin, gentamicin, ampicillin/sulbactam, and doxycycline.

India is annually producing 5.3 million metricton of meat i.e. the 5 th largest in the World (DAHD, 2017) with the world's largest livestock population which plays an important role in rural economy and livelihood. It produces 21% of global chicken meat production annually. The poultry industry is a high growing vertically integrated industry in India and as well as in the state of West Bengal. West Bengal is the 2 nd largest contributor with 640 thousand metric ton meat production of which chicken meat International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 9 Number 7 (2020) Journal homepage: http://www.ijcmas.com

In the current study prevalence and characterization of extended spectrum beta-lactamase(ESBL) production of Escherichia coli was detected from broilers of West Bengal, India. A total of 248 cloacal swabs samples were examined for E.coli by standard bacteriological techniques. E.coli isolates were serotyped and also screened for extended spectrum beta-lactamase by double disc diffusion assay and by PCR detection of blaCTX-M, blaSHV, and blaTEM genes. The antimicrobial sensitivity profile of the isolated E.coli was demonstrated against 13 antimicrobial agents. One hundred thirty one (52.82%) E.coli were identified and the isolates belonged to 12 different serogroups viz. O2, O20, O35, O83, O87, O88, O116, O117, O119, O128, O135, O141 and untyped isolates. Thirty seven (28.24%) of E.coli were detected as ESBL-positive by double disc diffusion assay. Among the studied genes, blaCTX-M gene was detected in 32 (24.43%), blaSHV gene in 3 (2.29 %) isolates and blaTEM gene in 2 (1.53%) isolates. Highest antimicrobial susceptibility of E.coli isolates was observed to chloramphenicol (69.36 %) and levofloxacin (62.9 %). Highest degree of resistance was observed to cefepime (95.12%) followed by nalidixic acid (88.88%), co-trimoxazole (83.33%), ampicillin (79.03%), cefotaxime (78.46%), tetracycline (72.58%) and ciprofloxacin (62.90%). The present study revealed that broilers were reserviors of ESBL producing E.coli and resistant to many classes of antimicrobials.

Two biovars of Salmonella enterica subsp. enterica biovar Gallinarum and Pullorum, causing fowl typhoid (FT) and pullorum disease of poultry, respectively are economically important in poultry industry. Eradication in commercial poultry in some parts of the world was achieved through improved surveillance and culling. However, threats of FT outbreaks are not eliminated in commercial and backyard poultry of developing countries (Barrow and Freitas Neto, 2011). Accurate diagnosis of the pathogen is prerequisite for effective adaptation of control measures. Flock history, mortality, clinical signs and post mortem lesions are suggestive of FT infection but isolation and biochemical identification of the organism still remain ̳gold standard‘ method of confirmation (OIE, 2018). The conventional methods of biochemical identification are laborious, and time consuming. Oand H-antigen specific antisera are used commonly for slide and tube agglutination tests to identify Salmonella serovars. Therefore, rapid detection technique of the major Salmonella serovars is utmost necessary. DNA based identification of Salmonella serovars Gallinarum and Pullorum International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 8 Number 06 (2019) Journal homepage: http://www.ijcmas.com

Background: Milk is considered as complete food and an important part of human diet throughout the world including India. Bacterial contamination of milk such as Escherichia coli due to unhygienic condition and poor udder health can cause infections, especially in infants and elders or in immunocompromised persons. Possession of antimicrobial resistance genes by commensal bacteria present in milk makes the issue more serious. Aim: The study was aimed to isolate and characterize extended-spectrum beta-lactamase (ESBL)-producing E. coli from milk samples collected from different parts of West Bengal, India, to assess the potential risk associated with the food. Materials and Methods: Around 182 milk samples were collected from apparently healthy cows reared by organized dairy farms in West Bengal. E. coli was isolated from collected samples as per standard methods followed by serotyping. The detection of ESBL-producing E. coli was done both phenotypically and genotypically by detecting the presence of blaCTX-M gene. Antibiogram of the ESBL-positive isolates was done using common 12 antibiotics by disc diffusion method. Results: A total of 22 (12.1%) samples were found to be positive for E. coli in this study. Different serotypes such as O11, O20, O22, O34, O35, O128, O149, and UT were isolated from the collected samples. 12 (54.5%) E. coli strains showed the capability of producing ESBL, both phenotypically and genotypically with the presence of blaCTX-M gene. Antibiogram of these ESBL-positive isolates revealed the drugs such as colistin (100%), levofloxacin (83.33%), and imipenem (66.67%) to be highly sensitive against this pathogen but drugs such as cefotaxime (100%), ceftazidime (91.67%), amoxicillin/clavulanic acid (83.33%), tetracycline (75.00%), and gentamicin (58.33%) to be very much resistant. Conclusion: More than 50% of the E. coli strains prevalent in the bovine milk samples were positive for ESBL production and are resistant to most of the common antimicrobials which may be alarming for human health.

Fatty liver haemorrhagic syndrome (FLHS) in poultry is a non-infectious metabolic disease of unknown aetiology. An outbreak of fatty liver haemorrhagic syndrome (FLHS) was detected at post mortem from 42-week old caged Vencobb broiler breeders in a commercial farm (n =1200) of Bankura, West Bengal, India. The affected flock was showing severe loss of production and sudden death in peak summer month of June with 4% mortality in a week. Post mortem of the dead birds were performed and bacteriological examination were done as per standard procedure. At necropsy, dead birds had pale combs and wattles with significant liver lesions. The liver was enlarged, friable and greasy, yellowish brown with firm fatty layer deposits and haematoma was noticed in abdominal cavity. Escherichia coli was isolated in bacteriological examination of liver and spleen. Multiple factors like high dietary energy, stress of production and high ambient temperature may have precipitated the FLHS in broiler breeder hens.

Preparation of live attenuated freeze dried Pigeon Paramyxo Virus-1 (PPMV-1) vaccine selection of proper cryoprotectant which could preserve and protect the virus against injury due to reduction of temperature at the time of lyophilization, was very necessary. PPMV-1 locally isolated lentogenic strain was selected as seed virus which was properly attenuated by serially passaging in 9 to 10 days old embryonated specific pathogen free (SPF) fowl eggs and produced master seed virus and working seed virus. Five per cent lactalbumin hydrolysate with 10% sucrose (LAS) and 1% polyvinyl pyrrolidone (PVP) were used as two different cryoprotectant for lyophilization of working seed virus. After lyophilization value of haemagglutination (HA) titer and embryo infective dose fifty per ml (EID50/ml) were changed and concluded that both LAS and PVP could be used as suitable cryoprotectant where LAS was better than PVP which was first time done in the whole world.

Aim: The aim was to characterize Salmonella enterica serovar Gallinarum isolated from backyard poultry by polymerase chain reaction (PCR) detection of virulence genes invasion (invA) and Salmonella plasmid virulence C (spvC). Materials and Methods: Two strains of Salmonella serovar Gallinarum isolates used in this study were obtained from an outbreak of fowl typhoid in backyard Vanaraja fowl. PCR technique was used for detection of invA and spvC genes using standard methodology. The invA PCR product from one representative isolate was sequenced and compared with other related Salmonella serovars in GenBank data. Results: Salmonella Gallinarum produced expected amplicons of invA and spvC gene products. Nucleotide sequence of 285 bp invA gene was deposited in GenBank with accession no. KX788214. Sequence analysis of invA gene was found conserved in Salmonella serovars and demonstrated 100% homology with closely related serovars of Salmonella. Conclusion: Invasion gene (invA) was found to be highly conserved in Salmonella Gallinarum and highly similar with closely related serovars. The isolates also contained plasmid-mediated spvC gene indicating possession of virulence plasmid.

the present study was undertaken to detect the incidence of extended-spectrum-cephalosporinase gene possessing-Escherichia coli, its co-resistance pattern against other antimicrobials, and the clonal relationship of the isolates in healthy kuroiler birds.a total of 80 cloacal swabs from kuroilers were collected randomly from West Bengal, India.the use of costly antimicrobials (cephalosporins) was not practiced by farmers.Escherichia coli was isolated and identified by standard biochemical tests and 16SrRNA-PCR.All the E. coli isolates, including controls, were subjected to PCr for detection of bla CTX-M , bla TEM, bla SHV, and bla CMY-2 genes.By comparing the raPD-banding pattern, the phylogenetic relationship among the isolates was established.all the isolates were tested for phenotypical resistance against other antibiotics.In total, 60 isolates were identified as E. coli from the kuroilers studied (n = 80).among them, 12 (20%) isolates possessed one of the studied extended-spectrum cephalosporinase genes.among the studied genes, bla TEM and bla SHV were detected in 6 (10%) and 12 (20%) E. coli isolates, respectively.None of the E. coli isolates possessed bla CTX-M and bla CMY-2.In phylogenetic analysis, the strains isolated from same localities with similar genetic profile were grouped into the same cluster.resistance of extended-spectrum cephalosporinase gene possessing E. coli isolates was observed most frequently against ampicillin/cloxacillin, co-trimoxazole, amoxyclav, piperacillin, ceftriaxone, and tetracycline.Kuroiler birds with no cephalosporin usage profile may act as a reservoir of extended-spectrum cephalosporinase gene possessing E. coli.This is the first systematic study in kuroilers, to raise the awareness of consumers regarding the possibility of transmission of antimicrobial resistant E. coli from them.

Aim: With the aim of revealing the epidemiological intricacies of bluetongue (BT) in the southern part of West Bengal state, the present study was undertaken to assess seroprevalence of BT along with identification of the vector of the disease, i.e., Culicoides midges available in the region in their breeding season with conducive environmental factors, if any. Materials and Methods: A total of 1509 (sheep-504, goat-1005) samples were collected from three different agroclimatic zones of South Bengal viz. new alluvial, red laterite and coastal saline. To detect anti-BT antibodies in the collected serum samples, indirect-enzyme-linked immunosorbent assay (i-ELISA) was performed. Culicoides midges were collected from those agro-climatic zones of South Bengal for species identification. The meteorological parameters, viz. temperature (maximum and minimum), rainfall and relative humidity of three agro-climatic zones of South Bengal were analyzed for the months of July to December during 2010-2013. Results: The overall seropositivity was 33.13% and 30.24% in sheep and goat, respectively as assessed by i-ELISA. In South Bengal, the predominant species of Culicoides found were Culicoides schultzei, Culicoides palpifer and Culicoides definitus. Conclusion: Since virus transmitting species of Culicoides midges could be detected in South Bengal, besides high seropositivity in ruminants, the possibility of circulating BT virus in South Bengal is quite imminent.

Aim: This study was carried out to assess the presence of anti-bluetongue (BT) antibodies in sheep, goat and cattle of different agro-climatic zones of Jharkhand. Materials and Methods: Serum samples were collected from apparently healthy as well as suspected sheep, goat and cattle from different districts of Jharkhand covering different agro-climatic zones. Serum samples were screened by indirect enzyme linked immunosorbent assay (iELISA) for detecting anti-BT antibodies. Results: Out of a total of 480 animal serum samples (sheep-190, goats-210 and cattle-80) screened, 83 (43.68%) of sheep, 91 (43.33%) of goat and 46 (57.50%) of cattle sera were found positive. The % positivity ranged between 41% and 51% in different agro-climatic zones. The results showed slight higher seroprevalence, although not significantly, in cattle than sheep and goats in different agro-climatic zones of Jharkhand. Conclusions: The above data indicate widespread prevalence of BT virus antibodies in studied areas. The incidence of BT is not detected officially, so far. The present seroprevalence status of BT in Jharkhand indicates presence of BT infection in the state for the first time.

This study was undertaken to observe the prevalence, serogroup, avian pathogenic Escherichia coli (APEC)-associated virulence gene, randomly amplified polymorphic DNA (RAPD) pattern, and antibiotic resistance genes of E. coli in backyard layers and their environment in India. From the 360 samples of healthy layers and their environment, 272 (75.5%) E. coli were isolated. The majority (28.67%) of them were untypeable. Among the studied virulence genes (papC, tsh, iucC, astA), 52 (14.32%) isolates were found to possess astA, including the isolates from the drinking water of the birds (4/272, 1.47%). These strains belonged to 18 different serogroups. Most of the isolates were typeable by RAPD and they produced different patterns. Phenotypic resistance of the isolates was most frequently observed to erythromycin (95.83%), chloramphenicol (87.52%), and cotrimoxazole (78.26%). None of the isolates was found to possess extended-spectrum beta-lactamases (bla(TEM), bla(SHV), bla(CTX-M) or quinolone resistance (qnrA) genes by PCR. The present study was the first attempt in India to assess APEC distribution in backyard poultry production.