Effect of feeding Bengal gram residual forage-based pelleted total mixed ration on growth performance, nutrient availability, carcass traits and composition in finisher Barbari kids.

The study aimed to assess the impact of feeding Bengal gram residual forage-based pelleted Total Mixed Ration (TMR) with varying concentrate (C) to roughage (R) ratios on feed intake, nutrient utilization, growth, and carcass characteristics in Barbari kids. Sixteen weaned male Barbari kids (av. age, 233 ± 11 days; weight, 13.86 ± 0.76 kg) were divided into two groups (T1 and T2), each receiving different pelleted diets (TMR) with distinct concentrate to roughage ratios (T1 with 60:40; T2 with 40:60). The kids were fed for 133 days, and a digestion trial was conducted at the end of the study. After completion, all kids were slaughtered. Although, kids under T1 consumed higher (P < 0.001) amount of dry matter, and crude protein compared to T2, which was due to a higher concentrate to roughage ratio in T1. But, the average daily body weight gain (ADG) of finisher kids was 88.53, and 79.83 g/d/kid in T1 and T2, respectively; however, the difference was non-significant. Digestibility of organic matter, crude protein, and total carbohydrate was also greater in T1 compared to T2. Total digestible nutrients intake was higher (P < 0.001) in T1; similarly intake of digestible energy, and metabolizable energy were significantly increased (P < 0.01) in T1 compared to T2. Concentrations of volatile fatty acids and NH3-nitrogen were also enhanced (P < 0.05) in T1 compared to T2. We observed similar carcass weight, and dressing percentage in both groups, and carcass composition remained unaffected. The pelleted diet containing greater ratio of concentrate: roughage (60:40) had no additional benefits in terms of ADG, and carcass traits in finisher kids. Therefore, it may be concluded that the Bengal gram residual forage-based pelleted TMR diet containing C40: R60 (TDN 57.13%, DCP 7.64%, ME 9.11MJ/kg feed) is suitable for optimizing growth performance with desirable carcass traits, and meat composition in finisher Barbari kids reared under the intensive system.

Rapid detection of Actinobacillus pleuropneumoniae targeting the apxIVA gene for diagnosis of contagious porcine pleuropneumonia in pigs by polymerase spiral reaction.

Actinobacillus pleuropneumoniae is the primary aetiological agent of contagious porcine pleuropneumonia associated with serious economic impact on pig husbandry worldwide. Diagnosis of the disease by existing techniques including isolation and identification of bacteria followed by serotyping, serological techniques, conventional PCR, real-time PCR and LAMP assays are cumbersome, time-consuming, costly and not suitable for rapid field application. A novel isothermal polymerase chain reaction (PSR) technique is standardized for all the reagents, incubation time and incubation temperature against A. pleuropneumoniae. The sensitivity of the assay was determined against various dilutions of purified DNA and total bacterial count. The specificity of the assay was determined against 11 closely related bacterial isolates. The relative sensitivity and specificity were compared with bacterial isolation, conventional PCR and real-time PCR assays. The PSR assay for specific detection was standardized at 64°C for 30 min of incubation in a water bath. The result was visible by the naked eye after centrifugation of the reaction mixture or after incorporation of SYBR Green dye as yellowish-green fluorescence. The technique was found to be 100% specific and equally sensitive with real-time PCR and 10 times more sensitive than conventional PCR. The PSR assay could be applicable in the detection of the organisms in porcine nasal swabs spiked with A. pleuropneumoniae. This is the first-ever report on the development of PSR for specific detection of A. pleuropneumoniae and can be applied for early diagnosis at the field level.

Comparative occurrence of ESBL/AmpC beta-lactamase-producing Escherichia coli and Salmonella in contract farm and backyard broilers.

The present study was conducted to detect the occurrence of beta-lactamase and biofilm-producing Escherchia coli and Salmonella in apparently healthy broiler birds reared in household and contract poultry farms. In total, 150 cloacal swabs were collected from apparently healthy broiler birds of various age groups reared in backyard (n = 100) and contract farms (n = 50) in West Bengal (India). The isolation rate of ESBL producers was significantly more (P < 0·05) reared in contract poultry farms than those reared in backyard. Majority of the E. coli isolates possessed blaCTX-M followed by blaSHV and blaTEM . Majority of the Salmonella strains possessed blaTEM followed by blaCTX-M , and no blaSHV was detected. The selected sequences of the PCR products were found cognate with blaCTX-M-1 , blaCTX-M-2 , blaCTX-M-9 , blaCTX-M-14 , blaCTX-M-15 , blaSHV-12 and blaTEM-52 . The study detected 46·8% of E. coli and 42·5% of Salmonella strains as biofilm producers. Beta-lactamase-producing Enterobacteriaceae strains showed resistance against tetracycline, chloramphenicol, doxycycline, co-trimoxazole and ampicillin and sensitivity to imipenem-EDTA, colistin and gentamicin. The study revealed the partial clonal relationship of ESBL sequences possessed by the poultry isolates of the present study and local clinical isolates available in the database. The study made consumer awareness about careful handling of live birds or poultry meat to avoid the zoonotic transmission of antimicrobial-resistant Enterobacteriaceae.

Effect of supplementation of phytogenic feed additives on intake, in vitro fermentation, growth performance and carcass traits in weaned Barbari kids reared under intensive feeding.

Twenty-four weaned male Barbari kids (age 144.67 days; weight 11.99 ± 0.49 kg) were divided equally into three groups (T1, T2, and T3) in order to investigate the effect of supplementation of phytogenic feed additives (herbal mixture) in the complete pelleted feed on growth performance, in vitro rumen fermentation and carcass quality in kids reared under stall-fed condition. Treatment groups were as follows: T1, concentrate mixture (40%) plus arhar (Cajanus cajan) straw (60%) in total mixed ration (TMR) form fed ad libitum; T2, T1 diet in complete feed pellets form fed ad libitum; and T3, T1 diet in complete feed pellets form supplemented with herbal mixture (Tulsi/Haldi/Amla/Arni; ratio 1:1:1:1 on DM basis) at 0.5% in complete feed fed ad libitum. The experimental kids in each group were allowed for feeding for 8 months by following the respective feeding schedule. Rumen fermentation pattern under in vitro system was also studied using the same three diets as substrates. After 240 days of feeding, all goats were slaughtered following standard protocol. Total body weight gain (kg) and average daily gain (ADG, g/day/kid) were 18.57, 22.26, and 23.06 kg, and 79.91, 101.49, and 100.18 g in T1, T2, and T3 treatments, respectively. Pelleting of TMR (T2) and supplementation of herbal mixture in pelleted feed (T3) increased (P < 0.001) average daily weight gain in Barbari kids compared to T1 (TMR). Average dry matter intake (DMI, g/day/kid) during growth trial was greater (P < 0.05) in T3 (1079.17) than T1 (849.76) and T2 (968.76). Feed conversion efficiency was 8.92, 9.48, and 8.68% in T1, T2, and T3, respectively. The difference was statistically non-significant among the treatments. Supplementation of herbal mixture in the complete pelleted substrate had adjunct effect on improvement of TCA-precipitable-N and total VFAs in the incubation medium under in vitro system. Carcass weight (kg) tended to increase in finisher kids under T2 (16.58) and T3 (16.70) than T1 (14.61), but the variation was non-significant. The dressing percentage was similar among three treatments. Similarly, the muscle protein, fat, and cholesterol contents remained unaffected by different dietary treatments. Therefore, it may be concluded that densification of feeds in the form of complete pelleted feed and further supplementation with potential phytogenic feed additives increased total DMI and ADG and tended to enhance meat production potential in finisher Barbari kids without changing the meat chemical composition.

Multidrug resistant staphylococci isolated from pigs with exudative epidermitis in North eastern Region of India.

Exudative epidermatitis or greasy pig disease (GPD) is a contagious disease of pig and endemic worldwide caused by toxigenic strains under genus Staphylococcus. The present study reported an outbreak of GPD in Champhai district of Mizoram adjoining to the southern border of Myanmar. A total of 60 samples were collected from 22 clinically affected animals and processed for isolation and identification of Staphylococcus spp. All the isolates were subjected to antimicrobial sensitivity assay, biofilm production assay and detection of virulence genes, biofilm genes and mec genes followed by cloning and sequencing for phylogenetic analysis. A total of 44 staphylococci belonged to four species (S. sciuri, S. aureus,S. lentus, and S. hyicus) were isolated. Majority of the isolates were multidrug resistant with maximum resistance against ampicillin, penicillin including vancomycin. None of the S. hyicus isolates was methicillin resistant (MRSH) but 66·67% isolates were MRSA. By PCR, mecA gene was detected in S. aureus (n = 2), S. sciuri (n = 4) and S. lentus (n = 3). Biofilm associated gene icaD was detected in S. aureus (n = 3), S. sciuri (n = 5), S. hyicus (n = 4) and S. lentus (n = 6). The exfoliative toxin genes (ehxB, shetA and tsst1) were detected in S. hyicus (n = 3) and S. aureus (n = 1) isolates. All the isolates were closely related with the isolates from pigs of China, Germany, Japan and USA. The pathogens might be transmitted through illegal migration of pigs from Myanmar to India.

Impact of environmental factors on physiological adaptability, thermo-tolerance indices, and productivity in Jersey crossbred cows.

Environmental stressors such as temperature, humidity, rainfall, photoperiod, etc. under tropical conditions are some of the important limiting factors for optimum expression of production and reproduction in dairy cows. The present study evaluated changes in cardinal physiological responses, heat tolerance indices, and milk yield in Jersey crossbred cows due to abiotic stress. Climograph was developed using daily maximum and minimum temperatures, humidity, and rainfall at livestock farm over a period of 15 years. Seasons significantly (P < 0.01) influenced cardinal physiological responses and thermal adaptability of Jersey crossbred cows. Diurnal environmental variations between morning and afternoon of the shed caused increase in body temperature, pulse, and respiration rate by 0.59-0.91%, 4.95-7.18%, and 9.73-32.37%, respectively. Thermal adaptability of cows showed significant (P < 0.01) higher deviations from perfect state of adaptability during summer, monsoon, and post-monsoon as compared to winter and spring seasons. The responsiveness and ranking of cows were dissimilar among 3 heat tolerance indices, viz., Iberia heat tolerance index, Benezara's coefficient of adaptability, and Dairy Search Index (DSI). Enhanced respiration rate was the most important observable bio-indicator to poor thermal adaptability. Good thermo-tolerant cows had 8.86% higher (P < 0.05) monthly milk yield per cow than poorly adapted ones. Findings of this study suggest that deviations in cardinal physiological responses across the seasons are very reliable and simple estimates for evaluating levels of thermo-tolerance in crossbred cows.

Effect of heat stress amelioration through open-ridge ventilated thatched roof housing on production and reproduction performance of crossbred Jersey cows.

The present study was conducted to assess the impacts of loose house roofing designs and materials on modulation of micro-climate of the cow shed and its influences on production and reproduction performance of lactating crossbred Jersey cows. The control group (T0) was kept in a traditional single-slope asbestos roof shed and treatment group (T1) at a specially designed open-ridge ventilated, double-slope thatch roof shed. Lactation stage, average daily milk yield, and average parity at the beginning of the experiment were kept similar between T0 and T1 cows. The specifically designed T1 shed had significantly (P < 0.05) lower temperature, humidity, and thermal humidity index as compared to the traditional shed (T0). The T1 shed was cooler and more comfortable than T0. Housing significantly (P < 0.05) influenced cardinal physiological parameters of resident cows. Body temperature, pulse rate, respiration rate, and discomfort index were significantly (P < 0.05) higher in T0 as compared to those in T1 cows. Improved housing significantly (P < 0.01) influenced milk yield, composition, quality, and udder health of cows. Daily milk yield depending on parity, monthly milk yield, milk fat, SNF, and protein were significantly (P < 0.01) higher in T1 than those in T0 cows. The milk somatic cell count, modified California mastitis test score, electrical conductivity, and pH of milk samples of T0 cows were significantly (P < 0.0) higher than those of T1. Effect of housing type, parity, and interaction of housing × parity did not cause any significant (P > 0.05) differences in reproductive traits such as service period, break period, calving interval, pregnancy rate, and other cow fertility parameters. Post-partum body condition score and body weight loss did not differ significantly (P > 0.05) between the two groups; however, more loss occurred in T0 than T1. The estimated net profit from housing of cows (N = 20) in a comfortable shed was 2200 kg milk per annum, i.e., 110 kg/cow/annum. The study concluded that a central open-ridge ventilated, double-slope thatched roof design created a favorable micro-environment and it significantly affected cows' comfort index; thermo-comfortable housing significantly reduced the heat stress-oriented drop in milk yield and decline in milk composition, and improved udder health and other milk production traits; however, its impact was non-significant on reproduction and fertility traits in Jersey crossbred cows.

Multi-drug resistant, biofilm-producing high-risk clonal lineage of Klebsiella in companion and household animals.

Antimicrobial resistance is a global emergency which needs one health approach to address. The present study was conducted to detect the prevalence of beta-lactamase and biofilm-producing Klebsiella strains in rectal swabs (n = 624) collected from healthy dogs, cats, sheep and goats reared as companion or household animals in India. The dogs and cats were frequently exposed to third- or fourth-generation cephalosporins for therapy. The sheep and goats were occasionally exposed to antibiotics and had environmental exposure. Phenotypical ESBL (n = 93) and ACBL (n = 88)-producing Klebsiella were isolated significantly more (P < 0·05) from companion animals than household animals. Majority of the Klebsiella possessed blaCTX-M-15 . The sequences blaCTX-M-15.2 , blaCTX-M-197 and blaCTX-M-225 are reported first time from the companion animals. All ACBL-producing isolates possessed blaAmpC . The present study detected 65·8% of Klebsiella strains as biofilm producers possessing the studied biofilm associated genes. The isolates showed phenotypical resistance against chloramphenicol, tetracycline, doxycycline, co-trimoxazole, ampicillin, cefotaxime/clavulanic acid. The present study showed that companion and household animals (dogs, cats, sheep, goats) may act as a carrier of ESBL/biofilm-producing, multi-drug resistant, high-risk clonal lineage of Klebsiella.

Extended spectrum beta-lactamase producing Shiga-toxin producing-Escherichia coli in piglets, humans and water sources in North East region of India.

Extended spectrum ß-lactamases (ESBL) producing Shiga toxin producing E. coli (STEC) are posing a constant threat to public health throughout the world leading to serious infections and raising key therapeutic issues. A total of 219 fecal samples were collected from piglets with diarrheoa, pig farmers and water sources in North East India; and were processed for isolation of Escherichia coli. The isolates were screened for antimicrobial resistance and suspected isolates for ESBLs production by double-disk synergy test (DDST). Escherichia coli isolates positive for DDST were subjected for detection of selected ESBL/beta-lactamase genes and virulence associated genes by PCR. By DDST, 337 (67·94%) E. coli isolates were detected as ESBLs producer, of which 211 (66·98%), 117 (70·91%) and 9 (56·25%) isolates were from piglets, humans and water sources respectively. A total of 64 (12·90%) isolates were recorded as STEC, of which 48 (9·68%), 6 (1·21%) and 10 (2·02%) were from human, piglets and water respectively. Majority of the STEC isolates (64·06%) possessed multiple virulence genes, of which 59·38% also harboured ESBL/beta-lactamase genes with 32·81% STEC isolates being positive for multiple ESBL/beta-lactamase genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Multidrug resistant (MDR) enteric bacteria are global concern. Association of MDR traits in STEC isolates are another rising issue in human and animal health perspective. The interaction of such organisms among the human, domestic animals and adjoining water sources require to be analysed systematically. The present study exhibited the possible transmission of MDR-STEC among the human, domestic animals and water sources in the North eastern states of India. To the best of our knowledge, this is the first report of such kind in India.

Characterization of beta-lactamase and biofilm producing Enterobacteriaceae isolated from organized and backyard farm ducks.

This study was undertaken to detect the occurrence of beta-lactamase and biofilm producing Enterobacteriaceae in healthy ducks. A total 202 cloacal swabs were collected from ducks kept in organized (n = 92) and backyard (n = 110) farms in West Bengal (India). The ducks had no history of antibiotic intake. Among the 87 phenotypically beta-lactamase producing Escherichia coli, 19 (17·43%), 6 (5·05%) and 15 (13·76%) isolates possessed blaTEM , blaSHV and blaCTX-M respectively. Whereas, 5 (38·46%) Salmonella isolates were found to harbour blaCTX-M . In K. pneumoniae 10 (33·33%), 3 (13·33%), 4 (13·33%) isolates possessed blaTEM , blaSHV and blaCTX-M respectively. The sequences of selected PCR products were found 98% cognate with blaCTX-M-9, blaSHV-12 and blaTEM-1 . Beta-lactamase producing E. coli isolates belonged to 14 different serogroups such as O1, O2, O3, O5, O7, O8, O35, O83, O84, O88, O119, O128, O145 and O157. Moreover, 87 E. coli (79·82%), six Samonella (46·15%) and 13 K. pneumoniae (43·33%) isolates were detected as AmpC producers possessing blaAmpC . Majority of E. coli (46·79%), Salmonella (46·15%) and K. pneumoniae (70%) isolates were detected as biofilm producers and possessed the associated genes (csgA, sdiA, rcsA, rpoS). Significantly higher occurrence of beta-lactamase and biofilm producing Enterobacteriaceae isolates was detected in backyard ducks than organized farms. SIGNIFICANCE AND IMPACT OF THE STUDY: Consumption of antibiotic through feed or during therapy is considered as potential reason for generation of antimicrobial resistant bacteria in birds. This study provides valuable evidence that exposure to contaminated environment may be an additional source for generation of antimicrobial resistant bacteria in backyard ducks. The backyard ducks are reared by marginal farmers in India who cannot offer antibiotics to them either through feed or during therapy due to high cost. The study also reveals a significant correlation between biofilm formation and possession of antimicrobial resistance genes in the bacterial isolates from the ducks.

Impact of extension interventions in improving livelihood of dairy farmers of Nadia district of West Bengal, India.

Livestock is a one of the major sources of livelihood for most of the small and marginal farmers in India, particularly for rural households who live in below poverty line. Extension interventions have long been seen as a key element for enabling farmers to obtain information and technologies that can improve their livelihoods. It is also recognized that extension is an important factor in promoting dairy development. Ex-post-facto cause to effect research design was applied in this study to trace out the impact of extension interventions in improving knowledge, attitude, adoption towards scientific dairy farming practices and improvement in milk production of dairy animal and income from dairying which will be resulted into improved livelihood of rural poor in Nadia district of West Bengal, India. Therefore, 60 dairy farmers of experimental villages who were considered as beneficiaries and 60 dairy farmers of control villages who were considered as non-beneficiaries were selected as sample for the study. It was found that beneficiaries had significantly higher score in all the five components of livelihood improvement with its all sub components, i.e., knowledge, attitude, adoption of scientific dairy farming practices, milk production per household per day and monthly income from dairying except disease control, and marketing component of adoption. Hence, it may be concluded that extension interventions had a significant impact on improving livelihood of rural dairy farmers in Nadia district of West Bengal, India.

Prevalence and molecular characterization of porcine Picobirnavirus in piglets of North East Region of India.

Picobirnaviruses (PBVs) have been recognized as one of the important causal viral agents of gastroenteritis in several animal species especially in young immunocompromised hosts. In this study, we report the prevalence and molecular epidemiology of porcine PBVs from North Eastern Hilly region of India. A total of 457 fecal samples from piglets were collected from local (n = 130) and cross (n = 327) breed piglets in different seasons for 2 years. All the samples were subjected to RNA-PAGE and RT-PCR analysis for detection of PBVs. A total of 4.59 and 11.15% samples were recorded as positive for PBVs by RNA-PAGE and RT-PCR, respectively. Rate of detection was higher from diarrhoeic animals (13.56%) compared to non-diarrhoeic (4.23%) animals. Higher prevalence rate was observed from unorganized farms (14.22%) compared to organized farms (8.0%) with slightly higher detection from cross breed (11.62%) compared to local breed (10.0%). Maximum cases of piglet diarrhea associated with PBVs were detected during summer (16.4%) and winter (14.39%) seasons compared to autumn (4.80%) and spring (6.45%). All the samples were positive for PBV genogroup I only. Based upon the sequence analysis, the isolates were unique and placed in separate clad and were not closely associated with any other Indian isolates of PBVs so far. Two isolates were closely related with one Chinese isolate recovered from sewage water. This is the first systematic study of prevalence of PBVs associated with piglet diarrhea.

A randomized study of very low-dose factor VIII prophylaxis in severe haemophilia - A success story from a resource limited country.

INTRODUCTION: Current factor prophylaxis strategy practised in developed countries is not feasible in resource constraint developing countries like India. AIM: The aim of this study was to investigate the efficacy and safety of very low-dose factor prophylaxis in India. METHODS: Children of 1-10 years of age with severe haemophilia were randomized to Prophylaxis group and Episodic (On demand) group. Children in prophylaxis group received very low-dose factor VIII (FVIII) concentrate, i.e. 10 units kg(-1) body weights on 2 days a week. Episodic group received factor concentrate in standard recommended doses. The study period was 11.5 months. RESULTS: In total 21 children were enrolled in this study, 11 assigned to prophylaxis and 10 to episodic group. Children on prophylaxis had 11 joint bleeds in comparison to 57 joint bleeds in episodic group. Mean number of haemarthrosis per patient per month were 0.08 (0.08 ± 0.13) in prophylaxis group compared to 0.48 (0.48 ± 0.34) in episodic group (P < 0.05). Total FVIII consumption was 87.51 and 56.32 units kg(-1) month(-1) in prophylaxis and episodic group respectively (P = ns). Overall median hospital emergency visits were 1 day in prophylaxis group and 9 days in episodic group (P ≤ 0.05). Median days of absenteeism from school were 25 days in episodic group and 3 days in prophylaxis group (P < 0.05). No significant complications were noted in prophylaxis group and compliance was 98%. CONCLUSION: To conclude, low-dose FVIII prophylaxis is cost effective, efficacious and a safe method of preventing joint bleeds and consequent joint damages.

Near fatal spontaneous intraperitoneal bleeding: A rare manifestation in a congenital factor X deficiency carrier.

Congenital factor X (FX) deficiency is a rare coagulation disorder of autosomal recessive inheritance, characterized by bleeding of variable severity. Bleeding severity generally correlates with the level of FX functional activity and severe bleeding usually occurs in moderate and severe deficiency, when FX coagulant activity is <5%. FX activity above 10% is infrequently associated with severe bleeding. Here we report the rare occurrence of life-threatening massive spontaneous intraperitoneal bleeding with hypovolemic shock, resulting from spontaneous rupture of an ovarian luteal cyst in a 25-year-old FX deficiency carrier woman, with a FX activity of 26%. She was managed successfully conservatively, with fresh frozen plasma and packed red blood cell transfusions and she showed gradual improvement. The case is being reported to discuss the diagnosis and management of this rare inherited coagulation disorder.

Inferior vena caval thrombosis: a rare complication of acute pancreatitis.

Inferior vena cava [IVC] thrombosis is a rare complication of acute pancreatitis. Here we report a case of IVC thrombosis with associated portal vein thrombosis complicating recurrent acute pancreatitis related to alcohol in a 31 year old male patient. We discuss the pathogenesis, clinical features and management of IVC thrombosis complicating acute and chronic pancreatitis in brief. The case is being reported for its rarity and early diagnosis as well as prompt treatment are essential for prevention of complications like pulmonary embolism.

Isolation, molecular characterization and antibiotic resistance of Shiga Toxin-Producing Escherichia coli (STEC) from buffalo in India.

In total, 363 Escherichia coli were isolated from 165 faecal samples of healthy buffaloes in West Bengal, India. Twenty-four of these isolates (6·61%) were found to carry at least one gene characteristic for Shiga toxin-producing Escherichia coli (STEC). These STEC strains belonged to 13 different O-serogroups. The stx1 gene was present in 23 (95·8%) of total STEC isolates, whereas 20 (83·3%) STEC isolates carried the gene stx2. Twelve strains of E. coli (50% of total STEC isolates) possessed enterohaemolysin (ehxA) gene in combination with others. Fourteen (58·33%) isolates found to possess saa gene. However, no E. coli was detected harbouring gene for intimin protein (eaeA). Of 23 stx1 -positive isolates, seven (30·43%) were positive for genes of the stx1C subtype. Of the 20 isolates with the stx2 gene, 25% (5/20) possessed stx2C and 10% (2/20) possessed stx2d gene. The phylogenetic analysis after RAPD of STEC strains revealed six major clusters. The isolated STEC strains were resistant most frequently to erythromycin (95·83%), cephalothin (62·5%), amikacin (54·17%), kanamycin (45·83%) and gentamicin (41·67%) group of antibiotics. No ESBL-producing (blaCTXM , blaTEM , blaSHV ) or quinolone resistance gene (qnrA) was detected in the STEC isolates.

PCR-Based Detection of Extended-Spectrum ß-Lactamases (bla CTX-M-1 and bla TEM ) in Escherichia coli, Salmonella spp. and Klebsiella pneumoniae Isolated from Pigs in North Eastern India (Mizoram).

Cephalosporins are major antimicrobials used to treat serious infections. However, their effectiveness is being compromised by the emergence of extended-spectrum ß-lactamases (ESBLs). A total of 138 enteric bacteria were isolated from 53 faecal samples of pigs collected from different districts of Mizoram, of which 102 (73.91 %) were Escherichia coli, 26 (18.84 %) were Salmonella spp. and 10 (7.25 %) were Klebsiella pneumoniae. Phenotypic confirmatory test (Double Discs Synergy Test) showed that 8 (5.80 %) E. coli isolates were ESBLs producer. PCR analysis confirmed that out of the eight isolate, 7 (5.07 %) harboured bla CTX-M-1 gene and/or bla TEM gene. Of the eight positive isolates, 7 (5.07 %) and 3 (2.17 %) were found to be positive for bla CTX-M-1 gene and bla TEM gene, respectively, of which 3 (2.17 %) isolates were positive for both the genes. Only 4 (2.90 %) E. coli isolates carried bla CTX-M-1 gene alone. Agarose gel electrophoresis showed that all the isolates were carrying plasmids ranging between 0.9 and ~30 kb. Out of the seven isolates positive for bla CTX-M-1 and/or bla TEM , 2 (1.84 %) isolates were confirmed for bla CTX-M-1 gene in their plasmid. Only one E. coli isolate was found to be positive for both the genes in its plasmid. The resistance plasmid could not be transferred to a recipient by in vitro horizontal gene transfer method.

Phenotypic & molecular characterization of AmpC ß-lactamases among Escherichia coli, Klebsiella spp. & Enterobacter spp. from five Indian Medical Centers.

BACKGROUND & OBJECTIVES: AmpC ß-lactamases which are often plasmid mediated hydrolyze all ß-lactam antibiotics except cefepime and carbapenems. We evaluated the presence of AmpC ß-lactamases among Enterobacteriaceae strains recovered prospectively from patients at five Indian tertiary care centres. METHODS: The study included 909 consecutive Gram-negative isolates recovered from clinically significant specimens during June 2007 - May 2008 as part of an ICMR-ESBL study. Among the study isolates, 312 were found to be cefoxitin resistant by disc diffusion test (DDT). Minimum inhibitory concentration (MIC) determination by E test was done against amikacin, levofloxacin, impinem, meropenem, ertapenem, tigecycline and piperacillin-tazobactam. Combined DDT using phenyl boronic acid as inhibitor with cefoxitin was used for phenotypic confirmation of AmpC phenotype. The common Amp C genotypes ACC, FOX, MOX, DHA, CIT and EBC were detected by multiplex PCR. RESULTS: Plasmid mediated Amp C phenotype was confirmed in 114 of the 312 (36.5%) cefoxitin resistant isolates with 255 (81.7%) showing multidrug resistance. Susceptibility to tigecycline was highest (99%) followed by imipenem, meropenem (97%), ertapenem (89%), amikacin (85%), and piperacillin-tazobactam (74.6%). Levofloxacin resistance was 82 per cent. ESBL co carriage was observed among 92 per cent of Amp C producers. Among 114 Amp C producers, 48 could be assigned a genotype, this included CIT- FOX (n = 25), EBC (n = 10), FOX (n = 4), CIT (n = 3), EBC-ACC (n = 2) and one each of DHA, EBC-DHA, FOX -DHA and FOX-EBC-DHA. INTERPRETATION & CONCLUSIONS: Overall, AmpC phenotypes were found in 12.5 per cent isolates, multidrug resistance and ESBL co-carriage among them was high suggesting plasmid mediated spread. The study results have implications in rational antimicrobial therapy and continued surveillance of mechanisms of resistance among nosocomial pathogens.

Type 2 lepra reaction as a cause of pyrexia of unknown origin.

Leprosy, a commonly encountered disease, can rarely present as a reactional state de novo with fever as the main presenting feature. Here we describe an uncommon presentation of leprosy [with type 2 lepra reaction] as pyrexia of unknown origin with prominent rheumatologic manifestations [acute polyarthritis], renal involvement and generalized lymphadenopathy with rare presentation of type 2 lepra reaction without the classic skin lesions of erythema nodosum leprosum, occurring in a treatment naive patient without prior history of leprosy.

Beneficial role of skim milk against drug-resistant Escherichia coli associated with pediatric diarrhoea.

Background: Antibiotic-resistance in E. coli is a global issue affecting humans especially the pediatric population. Antibiotic-resistant E. coli is a pathogen frequently pediatric population as well as healthy adults. Methods: This study aimed to examine the antibiotic resistance of E. coli causing pediatric diarrhea and its drug-resistant rates, its adhering abilities to cell line in vitro, and inhibition efficiency of a few selected chemical compounds. Clinical strains were isolated from both the healthy and infected pediatric population of Mizoram. Results: Adhesion is a significant pathogenic process during bacterial infections, which has been employed for pathotyping of DEC by comparing adhesion efficiency in both normal (CHO-k1) and cancer (HeLa) cell lines. E. coli adherent pathotypes were identified by both PCR assay and in-vitro cell adhesion assays; the study also evaluated the adhesion inhibition ability of human skimmed milk, gentamicin, and cephalexin in-vitro. Of all isolates, 20.05% of adherent DEC (EPEC, DAEC, and EIEC) and 11.39 % of non-adherent DEC (STEC and ETEC) were found to be associated with pediatric diarrhoea in Mizoram. Human skimmed milk has a high potential adhesion inhibition against EAEC (50.25/90.90 µg/mL), EPEC (53.42/259.70 µg/mL), and EIEC (59.13/30.30 µg/mL) in both cell lines in comparison with gentamicin and cephalexin. Conclusion: This study concludes that as a dietary supplement-human skimmed milk has high potential to prevent adhesion of DEC pathotypes in cells in-vitro thus in in-vivo.