A comparative study of two-dimensional and three-dimensional ultrasonography in evaluation of gastric affections in dogs

Research (Published online: 06-06-2015)

3.  A comparative study of two-dimensional and three-dimensional ultrasonography in evaluation of gastric affections in dogs - Madan Pal, Prem Singh, Rishi Tayal, Dinesh Dehmiwal, S. M. Behl, Sarvan Kumar and R. K. Chandolia

Veterinary World, 8(6): 707-712

   doi: 10.14202/vetworld.2015.707-712

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Abstract

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Aim: The objective of the study was to obtain and compare the two-dimensional (2D) and three-dimensional (3D) ultrasonographic images of pathological conditions of the stomach in dogs in clinical cases.

Materials and Methods: In our study, 12 clinical conditions of the stomach were recorded using ultrasonography. The ultrasound machine used for this study was 3D ultrasound machine (Nemio-XG: Toshiba, Japan) having four-dimensional volumetric transducer.

Results: Present study was done to compare 2D and 3D ultrasonographic images in different gastric affections in dogs. In case of uremic gastropathy due to inflammatory response, the wall of the stomach was 0.6 cm thick and hyperechoic and gastric folds were also hyperechoic indicative of gastritis. In second, third, and fourth case of gastritis the wall of the stomach was 0.7, 0.6, and 0.55 cm, respectively thick and hyperechoic. In fifth and sixth case of gastritis, inflammatory response due to ingestion of polythene and sand led to gastritis and ultrasonographically, the wall of the stomach was 0.6 cm and 0.7 cm thick, respectively, and hyperechoic. In case of gastric ulcer, ultrasonographically, there was a disruption of gastric mucosal layer. In cases of gastric dilatation, anechoic content indicating fluid was seen in stomach area and due to dilatation boundary of the stomach was not clear and the increase in the lumen of the stomach was observed. In case of foreign body, ultrasonographically the wall of the stomach was 0.55 cm thick and hyperechoic. In the middle of the stomach, multiple hyperechoic shadows of the foreign bodies i.e. leather and bunch of straw of grass were observed. In case of pyloric stenosis ultrasonographically, anechoic lumen of the pylorus surrounded by 0.5 cm hypoechoic thickened muscle. In some cases, 3D ultrasonography was not diagnostic i.e. gastric foreign bodies and gastric dilatation. These conditions were better visualized on the 2D sonogram.

Conclusion: The appearance of clinical conditions of the stomach such as gastritis and pyloric stenosis were more distinct on 3D ultrasonogram than 2D ultrasonogram. The 3D ultrasonography was not diagnostic in cases of gastric foreign bodies and gastric dilatation.

Keywords: dogs, gastritis, gastric dilatation, gastric foreign bodies, gastric ulcer, three-dimensional ultrasonogram.

Isolation and molecular characterization of Salmonella spp. from chevon and chicken meat collected from different districts of Chhattisgarh, India

Research (Published online: 06-06-2015)

2. Isolation and molecular characterization of Salmonella spp. from chevon and chicken meat collected from different districts of Chhattisgarh, India - V. K. Naik, S. Shakya, A. Patyal, N. E. Gade and Bhoomika

Veterinary World, 8(6): 702-706

   doi: 10.14202/vetworld.2015.702-706

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Abstract

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Aim: The aim was to assess the prevalence of Salmonella in raw chevon and chicken meat sold in the retail meat shops situated in and around Durg, Rajnandgaon, Dhamtari, Raipur, and Bilaspur districts of Chhattisgarh. Studies were also conducted to find out the antibiotic resistance in Salmonella isolates.

Materials and Methods: A total of 400 samples comprising of 200 chevon meat and 200 chicken meat samples were processed for isolation of Salmonella and all isolates were further confirmed on the basis of cultural and biochemical characters and by targeting invAgene of Salmonella. All Salmonella isolates were also examined for their antimicrobial drug susceptibility/resistance pattern against commonly used antibiotics.

Results: Out of 400 samples, the prevalence of Salmonella in chevon and chicken meat was found 9% and 7% respectively, with an overall prevalence of 8%. Polymerase chain reaction targeting invA gene of Salmonella showed positive result with 31 isolates. All 32Salmonella isolates were found to be highly sensitive to ciprofloxacin while 96.87%, 96.87% and 93.75% were sensitive to gentamicin, imipenem, and ceftazidime, respectively. 93.75% and 59.37% isolates were resistant to erythromycin and oxytetracycline, respectively. Out of 32, 14 isolates had multiple antibiotic resistance index equal to or more than 0.2.

Conclusion: Salmonella in chevon and chicken meat samples is prevailing in the areas of sampling due to poor hygienic conditions and also demonstrated the varied spectrum of antimicrobial resistance, including several multiple drug resistance phenotypes. Therefore, the present study emphasizes the need for continued  surveillance of zoonotic foodborne pathogens including antimicrobial-resistant variants throughout the food production chain.

Keywords: chevon, Chhattisgarh, chicken, isolation, molecular characterization, Salmonella. 

Isolation, identification, and characterization of Listeria spp. from various animal origin foods

Research (Published online: 06-06-2015)

1. Isolation, identification, and characterization of Listeria spp. from various animal origin foods - Deepti N. Nayak, C. V. Savalia, I. H. Kalyani, Rajeev Kumar and D. P. Kshirsagar

Veterinary World, 8(6): 695-701

   doi: 10.14202/vetworld.2015.695-701

Abstract

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Aim: The present study was undertaken with the prime objective of isolating and identifying Listeria spp. from various foods of animal origin sold at retail market outlets in the city of Navsari, Gujarat.

Materials and Methods: Total 200 samples comprising of milk, milk products, meat, and fish (50 each) collected aseptically from local market which were subjected first to pre-enrichment in half strength Fraser broth followed by enrichment in full strength Fraser broth and subsequent plating on PALCAM agar. The growth with the typical colony characteristics were further identified up to species level on the basis of their morphological and biochemical characteristics. Cultures identified as Listeria monocytogenes were further subjected toin vitro pathogenicity tests and detection of different virulence associated genes viz. actA, hlyA, and iap using polymerase chain reaction.

Results: Of the total 200 food samples of animal origin; 18 (9%) were found positive for Listeria spp. which were identified as Listeria seeligeri (6, 33.3%), Listeria innocua (5, 27.7%), Listeria welshimeri (4, 22.2%), and L. monocytogenes (3, 16.6%). The highest prevalence was observed in milk samples (8). Species wise, 6 isolates of L. seeligeri which included two each from cow milk, buffalo milk, and meat samples; 5 L. innocua isolates included four recovered from fish and one from meat sample; 4 L. welshimeri comprised of two isolates from ice cream and one each from buffalo milk and meat sample; and 3 isolates of L. monocytogenes recovered from milk (1 cow and 2 buffalo milk). All 3 L. monocytogenes isolates screened for the presence of virulence genes viz. actA, hlyA, and iapusing the specific primers revealed the presence of all the genes suggesting the possibility of danger of foodborne listeriosis among raw milk consumers.

Conclusion: Listeria spp. was isolated from 9% (18/200) of the animal origin food samples viz.; milk, milk products, meat, and fish with the highest prevalence in the milk samples. L. monocytogenes was isolated from 3 milk samples only. L. seeligeri was the predominant species isolated followed by L. innocua, L. welshimeri, and L. monocytogenes in this study. L. monocytogenes were found to carry virulence genes like actA, hly A, and iap genes suggesting the pathogenic potential of these isolates.

Keywords: animal origin foods, Listeria monocytogenes, Listeria spp., polymerase chain reaction, virulence genes.

Co-culture: A quick approach for isolation of street rabies virus in murine neuroblastoma cells

Research (Published online: 21-05-2015)

14. Co-culture: A quick approach for isolation of street rabies virus in murine neuroblastoma cells - A. Sasikalaveni, K. G. Tirumurugaan, S. Manoharan, G. Dhinakar Raj and K. Kumanan

Veterinary World, 8(5): 636-639

   doi: 10.14202/vetworld.2015.636-639

Abstract

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Background: Laboratory detection of rabies in most cases is based on detection of the antigen by fluorescent antibody test, however, in weak positive cases confirmative laboratory diagnosis depends on widely accepted mouse inoculation test. Cell lines like neuroblastoma have been used to isolate the virus with greater success not only to target for diagnosis, but also for molecular studies that determine the epidemiology of the circulating street rabies strains and in studies that look at the efficiency of the developed monoclonal antibodies to neutralize the different rabies strains. Due to the recent issues in obtaining ethical permission for mouse experimentation, and also the passages required in the cell lines to isolate the virus, we report herewith a co-culture protocol using the murine neuroblastoma (MNA) cells, which enable quicker isolation of street rabies virus with minimum passages.

Objective: This study is not to have an alternative diagnostic assay, but an approach to produce sufficient amount of rabies virus in minimum passages by a co-culture approach in MNA cells.

Materials and Methods: The MNA cells are co-cultured by topping the normal cells with infected cells every 48 h and the infectivity was followed up by performing direct fluorescent-antibody test.

Results: The co-culture approach results in 100% infectivity and hence the use of live mouse for experimentation could be avoided.

Conclusion: Co-culture method provides an alternative for the situations with limited sample volume and for the quicker isolation of virus which warrants the wild type strains without much modification.

Keywords: co-culture, isolation, fluorescent-antibody test, murine neuroblastoma, rapid tissue culture infection test.

Critical sources of bacterial contamination and adoption of standard sanitary protocol during semen collection and processing in Semen Station

Research (Published online: 21-05-2015)

13. Critical sources of bacterial contamination and adoption of standard sanitary protocol during semen collection and processing in Semen Station - Chandrahas Sannat, Ajit Nair, S. B. Sahu, S. A. Sahasrabudhe, Ashish Kumar, Amit Kumar Gupta and R. K. Shende

Veterinary World, 8(5): 631-635

   doi: 10.14202/vetworld.2015.631-635

Abstract

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Aim: The present investigation was conducted to locate the critical sources of bacterial contamination and to evaluate the standard sanitation protocol so as to improve the hygienic conditions during collection, evaluation, and processing of bull semen in the Semen Station.

Materials and Methods: The study compared two different hygienic procedures during the collection, evaluation and processing of semen in Central Semen Station, Anjora, Durg. Routinely used materials including artificial vagina (AV) inner liner, cone, semen collection tube, buffer, extender/diluter, straws; and the laboratory environment like processing lab, pass box and laminar air flow (LAF) cabinet of extender preparation lab, processing lab, sealing filling machine, and bacteriological lab were subjected to bacteriological examination in two phases of study using two different sanitary protocols. Bacterial load in above items/environment was measured using standard plate count method and expressed as colony forming unit (CFU).

Results: Bacterial load in a laboratory environment and AV equipments during two different sanitary protocol in present investigation differed highly significantly (p<0.001). Potential sources of bacterial contamination during semen collection and processing included laboratory environment like processing lab, pass box, and LAF cabinets; AV equipments, including AV Liner and cone. Bacterial load was reduced highly significantly (p<0.001) in AV liner (from 2.33±0.67 to 0.50±0.52), cone (from 4.16±1.20 to 1.91±0.55), and extender (from 1.33±0.38 to 0) after application of improved practices of packaging, handling, and sterilization in Phase II of study. Glasswares, buffers, and straws showed nil bacterial contamination in both the phases of study. With slight modification in fumigation protocol (formalin @600 ml/1000 ft3), bacterial load was significantly decreased (p<0.001) up to 0-6 CFU in processing lab (from 6.43±1.34 to 2.86±0.59), pass box (from 12.13±2.53 to 3.78±0.79), and nil bacterial load was reported in LAFs.
Conclusion: Appropriate and careful management considering critical points step by step starting right from collection of semen to their processing can significantly minimize bacterial contamination.
Keywords: bacterial contamination, critical sources, environment, laboratory equipments, Semen Station.

Serum metabolic and minerals profile in norgestomet primed postpartum anestrous surti buffaloes

Research (Published online: 21-05-2015)

12. Serum metabolic and minerals profile in norgestomet primed postpartum anestrous surti buffaloes - Sanjay C. Parmar, C. T. Khasatiya, J. K. Chaudhary, R. V. Patel and H. B. Dhamsaniya

Veterinary World, 8(5): 625-630

   doi: 10.14202/vetworld.2015.625-630

Abstract

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Aim: The study was undertaken to find out the serum metabolic and minerals profile in postpartum anestrous surti buffaloes treated with norgestomet ear implants alone and in combination with pregnant mare serum gonadotropin (PMSG).

Materials and Methods: The study was conducted on 18 postpartum anestrous Surti buffaloes divided into three groups of six animals each at random to conduct the experiment. The buffaloes in Group-I and Group-II were implanted with Crestar ear implant for 9 days together with 2 ml injection of Crestar solution given i/m on the day of the implant insertion. In Group-II, additionally 500 IU PMSG was given i/m on the day of implant removal, whereas the buffaloes in Group-III served as anestrous control group and received 5 ml Normal Saline i/m on day 0 and 9 as a placebo treatment.

Results: The overall serum total protein values did not differ significantly (p > 0.05) between time (days) intervals in any of the groups. The mean serum total cholesterol levels at 10th day and on the day of estrus were found significantly lower (p < 0.05) in the control group as compared to treatment Groups I and II. However, there was no significant difference (p > 0.05) at 10th day and on the day of estrus between treatment groups (T1 and T2). The overall mean serum cobalt, zinc, iron, and manganese values did not differ significantly (p > 0.05) between different time intervals among any of the groups, except copper which was significantly lower (p < 0.05) at 10th day in control group as compared to treatment groups.

Conclusion: Microelements cannot be synthesized in the body. Hence, it is concluded that the mineral mixture should be supplied daily in the animals ration to suffice the requirement of the trace elements. The mean serum metabolic and micro-minerals profiles in treatment and control groups revealed that overall mean serum total protein, cholesterol, copper, and zinc levels were apparently higher in treatment groups whereas, mean serum cobalt, iron, and manganese concentration had no consistent trend between treatment and control groups of Surti buffaloes.

Keywords: anestrous, buffaloes, cholesterol, micro-minerals, norgestomet, pregnant mare serum gonadotropin, protein.

Isolation and purification of beta-lactoglobulin from cow milk

Research (Published online: 15-05-2015)

11. Isolation and purification of beta-lactoglobulin from cow milk - Ranjit Aich, Subhasis Batabyal and Siddhartha Narayan Joardar

Veterinary World, 8(5): 621-624

   doi: 10.14202/vetworld.2015.621-624

Abstract

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Aim: The present study was undertaken to standardize a convenient method for isolation and purification of β-lactoglobulin (β-lg) from cow milk keeping its antigenicity intact, so that the purified β-lg can be used for detection of cow milk protein intolerance (CMPI).

Materials and Methods: Raw milk was collected from Gir breed of cattle reared in Haringhata Farm, West Bengal. Milk was then converted to skimmed milk by removing fat globules and casein protein was removed by acidification to pH 4.6 by adding 3 M HCl. β-lg was isolated by gel filtration chromatography using Sephacryl S-200 from the supernatant whey protein fraction. Further, β-lg was purified by anion-exchange chromatography in diethylaminoethyl-sepharose. Molecular weight of the purified cattle β-lg was determined by 15 percent one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was analyzed by gel documentation system using standard molecular weight marker.

Results: The molecular weight of the purified cattle β-lg was detected as 17.44 kDa. The isolated β-lg was almost in pure form as the molecular weight of purified β-lg monomer is 18kDa.

Conclusion: The study revealed a simple and suitable method for isolation of β-lg from whey protein in pure form which may be used for detection of CMPI.

Keywords: beta-lactoglobulin, ion-exchange chromatography, milk protein intolerance, sodium dodecyl sulfate polyacrylamide gel electrophoresis, whey protein.