Isolation, detection of virulence genes, antibiotic resistance genes, plasmid profile, and molecular typing among Vibrio parahaemolyticus isolated in Malaysian seawater from recreational beaches and fish

Research (Published online: 28-07-2019)

32. Isolation, detection of virulence genes, antibiotic resistance genes, plasmid profile, and molecular typing among Vibrio parahaemolyticus isolated in Malaysian seawater from recreational beaches and fish

Orooba Meteab Faja, Ali Abd Sharad, Khansa Mohammed Younis, Merriam Ghadhanfar Alwan, Basima Jasim Mohammed and Asmad Ahmad

Veterinary World, 12(7): 1140-1149

ABSTRACT

Background and Aim: Despite the importance of the global emergence of Vibrio parahaemolyticus infections worldwide, there has been scanty information on its occurrence in Malaysian seawaters and fish. This study aimed to determine the occurrence of V. parahaemolyticus isolates using polymerase chain reaction targeted at toxin operon gene, thermostable direct hemolysin (tdh), and tdh-related hemolysin genes and to determine antibiotic resistance pattern, genes, and plasmid profile of V. parahaemolyticus from Malaysian seawaters and fish.

Materials and Methods: Samples were collected from four recreational beaches in Malaysia (Port Klang; Bachok; Port Dickson; and Mersing). Thiosulfate-citrate-bile salts-sucrose (TCBS) agar and chromogenic Vibrio agar were used for isolation and identification. Colonies with yellow color on TCBS and green color on chromogenic vibrio (CV) agar were considered to be V. parahaemolyticus and they were subjected to biochemical tests. All V. parahaemolyticus isolates were further subjected to identification using seven specific gene markers.

Results: Seventy-three Vibrio isolates were recovered. Only one gene tdh> from seawater isolates of Vibrio has high virulence gene percentage (95.23%). Two genes alkaline serine protease (asp) and (tdh) had high percentage of virulence (83.87% and 80.64%, respectively) from fish. Comparatively, fish isolates have a higher virulence percentage compared to seawater isolates. Only gene streptomycin resistance B (strB) from seawater had 100% of the resistance genes. All isolates were multi-antibiotic resistant. Seventeen antibiotic resistance patterns were observed. The isolates had plasmids of varying sizes ranging from 2.7 kb to 42.4 kb. Dendrogram based on antibiotic resistance patterns of V. parahaemolyticus isolates discriminated the isolates into three clusters.

Conclusion: This study demonstrated the occurrence of pathogenic, multi-antibiotic-resistant V. parahaemolyticus strains in Malaysian coastal waters and fish, and this could constitute potential public health risks.

Keywords: antibiotic resistance genes, plasmid profile, Vibrio parahaemolyticus, virulence genes.

Retrospective study of the reproductive performance of Barb and Thoroughbred stallions in Algeria

Research (Published online: 26-07-2019)

31. Retrospective study of the reproductive performance of Barb and Thoroughbred stallions in Algeria

Nedjma Aouane, Abdelkrim Nasri, Mohamed Al Amine Bekara, Ahmed Khireddine Metref and Rachid Kaidi

Veterinary World, 12(7): 1132-1139

ABSTRACT

Background and Aim: The Barb horse occupies a prominent place in the history, culture, and equestrian traditions of the Maghreb (Algeria, Morocco, and Tunisia). Although many studies on the breed standard and morphometry have been published, there are no studies on its fertility and reproductive potential. Thus, this work aimed to study the fertility of Barb, Arabian Purebred, and Thoroughbred horses in Algeria.

Materials and Methods: A total of 168 stallions and 1202 mares at the Chaouchaoua Stud farm in Tiaret, Algeria, were included in the study. The reproductive performance during 1592 cycles over 10 consecutive mating seasons (2003-2012) was evaluated. Multivariate analysis with logistic regression was used to analyze the reproductive parameters such as the number of cycles operated, number of pregnant mares, pregnancy rate per cycle, seasonal pregnancy rate, and embryonic mortality rate, and to determine the influence of breed, stallion's age, and year of the study on reproduction.

Results: Statistical analysis showed that stallion breed was a significant influencing factor for the number of pregnant barren mares (Odds ratio [OR]=1.72; p=0.03; 95% confidence interval [CI]=1.05, 2.84) seasonal pregnancy rate (OR=1.40; p<0 .001="" 1.53="" 95="" additionally="" age="" barb="" ci="1.29," influence="" of="" performance="" regard="" reproductive="" s="" significant="" stallion.="" stallion="" superior="" that="" the="" thoroughbred="" to="" was="" with="">5 years on the number of pregnant foaling mares and seasonal pregnancy rate, significant interactions were observed between the stallion's breed and age, and the pregnancy rate per cycle: in the Barb breed, the pregnancy rate per cycle increased with the age of the stallion, while in the Thoroughbred, it decreased with age. Moreover, a significant effect of the year of the study on the pregnancy rate per cycle and number of pregnant foaling mares was observed. In contrast, the number of cycles and embryonic mortality rate were not influenced by the breed and age of the stallion, or the year of the study.

Conclusion: The Algerian Barb horse attained a similar level of fertility compared with that of the Arabian Purebred and Thoroughbred stallions depending on its age and reproductive performance.

Keywords: age, Barb, breeding performance, breed, stallion.

Serological detection of avian reovirus in different poultry flocks of Gazipur and Mymensingh districts of Bangladesh

Research (Published online: 26-07-2019)

30. Serological detection of avian reovirus in different poultry flocks of Gazipur and Mymensingh districts of Bangladesh

Syeda Farjana Neepa, Zobayda Farzana Haque, Abdullah Al Momen Sabuj, Md Alimul Islam and Sukumar Saha

Veterinary World, 12(7): 1126-1131

ABSTRACT

Background and Aim: Avian reovirus (ARV) is a constraint to poultry industry in Bangladesh as a cause of several diseases in chickens, especially in broiler. However, the actual status of the viral infection is not known because the large-scale study is not conducted in this country. Therefore, this study aimed to check the presence and distribution of ARV-specific antibody in respect to area, types of chickens (broiler breeder, broiler, and layer), vaccination status, and age of chickens in Gazipur and Mymensingh districts of Bangladesh.

Materials and Methods: A total of 276 chickens' blood samples were collected from two well-organized broiler breeder stock, seven broiler farms, and five layer farms located at two districts, namely Gazipur and Mymensingh of Bangladesh. Blood samples were collected from wing vein of the apparently healthy chickens using 3 ml of syringe and serum was harvested by keeping the syringe at room temperature in slanting position. The sera were transferred to the laboratory by maintaining the cool chain and further processing was performed by indirect enzyme-linked immunosorbent assay using ARV antibody test kit.

Results: The results of serological test revealed that an average of 39.5% seropositive against ARV was recorded in chickens of Gazipur and Mymensingh districts. Among these, chickens of Gazipur district had the highest seropositivity of 50.5% than Mymensingh (30.7%). With respect to vaccination status, the seropositivity of vaccinated chickens in both areas was 100% and non-vaccinated chickens was 50.5% in Gazipur and 30.7% in Mymensingh district, respectively. However, regarding age groups, the seropositivity was higher in the age of 4-6 weeks (64.5%).

Conclusion: The present serological findings showed a higher prevalence of ARV-specific antibodies in broiler birds. It indicates that the poultry industries of Bangladesh are contaminated with ARV which may naturally be transmitted to chickens either vertically or horizontally.

Keywords: avian reovirus, enzyme-linked immunosorbent assay, seropositive.

Abomasal displacement in neonatal dairy calves: Review of recent literature with special emphasis on abomasal torsion

Research (Published online: 25-07-2019)

29. Abomasal displacement in neonatal dairy calves: Review of recent literature with special emphasis on abomasal torsion

Zuhair Bani Ismail and Faisal Omoush

Veterinary World, 12(7): 1121-1125

ABSTRACT

Aim: This study aimed to describe the clinicopathological and necropsy findings in neonatal dairy calves affected with right torsion of the abomasum.

Materials and Methods: The history and findings of clinical examination, hematology and serum biochemical analyses and necropsy were described in six neonatal Holstein calves with a confirmed diagnosis of right torsion of the abomasum. Furthermore, a review of the literature was carried out using internet search engines such as PubMed and Google Scholar concerning abomasal displacement in calves. Only published papers in scientific and refereed journals were reviewed.

Results: Six neonatal Holstein calves (four females and two males) aged between 7 and 21 days were diagnosed with right torsion of the abomasum during necropsy. The calves were presented with peracute signs of anorexia, dehydration, abdominal pain, and abdominal distension. Hematology and serum biochemical analyses revealed hemoconcentration and azotemia, hyponatremia, hypochloremia, and hypokalemia.

Conclusion: Abomasal torsion in neonatal calves must be placed on the deferential diagnostic list of calves suffering from peracute signs of abdominal pain and distension. Although the underlying etiopathological factors are not fully known, correction of nutritional mismanagement is required to prevent the condition.

Keywords: abdominal surgery, abomasal diseases, Holstein calves, roughage feeding.

A diluent containing coconut water, fructose, and chicken egg yolk increases rooster sperm quality at 5°C

Research (Published online: 25-07-2019)

28. A diluent containing coconut water, fructose, and chicken egg yolk increases rooster sperm quality at 5°C

Siti Eliana Rochmi and Miyayu Soneta Sofyan

Veterinary World, 12(7): 1116-1120

ABSTRACT

Aim: The present study was conducted to evaluate the quality of rooster sperm at 5°C after treatment with a diluent containing coconut water, fructose, and chicken egg yolk and stored the semen sample at 5°C.

Materials and Methods: Ten semen samples from 10 healthy roosters were subjected to four different treatments. For the treatments, 0.2 ml fresh semen with a sperm concentration of 5.2X109 cell/ml was mixed with T0 (no diluent), T1 (0.34 ml coconut water and 6 μl fructose), T2 (0.274 ml coconut water, 0.12 ml egg yolk, and 6 μl fructose), and T3 (0.34 ml egg yolk and 6 μl fructose) solutions. Each treated solution was stored at 5°C and evaluated both macroscopically and microscopically. Macroscopically, semen volume, pH, and sperm concentration were evaluated. The microscopic sperm characteristics examined included total motility (i.e., rapid, medium, or slow), progressive and non-progressive motility, viability, and spermatozoa abnormalities noted at different storage times. The results showed that spermatozoa motility was under 40%.

Results: The results indicated that sperm viability significantly affected (p<0 .05="" 7="" abnormalities="" after="" day="" found="" highest="" lower="" mean="" of="" on="" significantly="" similarly="" solution="" span="" sperm="" spermatozoa="" storage="" t2="" the="" treatment="" value="" viability="" was="" were="" with="">

Conclusion: The addition of a diluent containing coconut water, egg yolk, and fructose helped in the better preservation spermatozoa motility, as well as viability for up to 7 days when the semen samples were stored at 5°C.

Keywords: chicken, coconut water, diluent, egg yolk, sperm quality.

Genetic analysis of NS5B gene from bovine viral diarrhea virus-infected cattle in Central and East Java, Indonesia

Research (Published online: 25-07-2019)

27. Genetic analysis of NS5B gene from bovine viral diarrhea virus-infected cattle in Central and East Java, Indonesia

S. H. Irianingsih, H. Wuryastuty, R. Wasito, H. Wibawa, F. S. Tjatur Rasa and B. Poermadjaja

Veterinary World, 12(7): 1108-1115

ABSTRACT

Background and Aim: A previous study divided Indonesian bovine viral diarrhea virus (BVDV)-1 into subgenotypes BVDV-1a to BVDV-1d based on the partial NS5B gene using strain Bega as reference for BVDV-1a. In fact, it is clustered into BVDV-1c with strain Bega-like Australia. BVDV genotyping has been done on isolates from Jakarta, West and Central Java, but East Java isolates have not been genotyped. This study aimed to analyze genetic variability and amino acid residues in the nucleotide-binding pocket of the NS5B gene from infected cattle.

Materials and Methods: Samples were obtained from the Sera Bank originating from active and passive surveillance of cattle that had been tested for BVDV antigen from 2013 to 2017. Detection of the p80 antibody and BVDV genotyping was carried out using ELISA and nested-multiplex-polymerase chain reaction (PCR), respectively. We defined 15 nested PCR products for partial sequencing of NS5B. Those field samples were selected from each location and year using proportional calculation as a representative sample. Homological and phylogenetic analyses of the partial NS5B gene were performed using BLAST and MEGA version 6.

Results: Based on the phylogenetic tree analysis using 360 nucleotides as the partial NS5B gene, Indonesian BVDV-1 isolates from Central and East Java were subdivided to BVDV-1a (n=9), BVDV-1b (n=1), and BVDV-1c (n=5). In the present study, the homology of BVDV subgenotype -1a, -1b, and -1c was compared to the BVDV GenBank data and found 90-93%, 93%, and 92-95% respectively with the average pairwise distance of 0.207. A point mutation was shown at R283K of all BVDV isolates based on the sequence of three amino acid residues R283, R285, and I287 in the nucleotide-binding pocket as a part of the encoded RNA-dependent RNA polymerase.

Conclusion: This study revealed the genetic variability of BVDV infecting cattle in Central Java and East Java, Indonesia, the subtypes BVDV-1a, BVDV-1b, BVDV-1c, and a point mutation at the R283K residue.

Keywords: bovine viral diarrhea virus, NS5B gene, phylogenetic analysis, point mutation, subgenotype.

The concentration of testosterone, pituitary adenylate cyclase-activating polypeptide, and protamine 1 in the serum of male chicken following administration of epididymis and testicular extracts and their combination

Research (Published online: 25-07-2019)

26. The concentration of testosterone, pituitary adenylate cyclase-activating polypeptide, and protamine 1 in the serum of male chicken following administration of epididymis and testicular extracts and their combination

Muslim Akmal, Gholib Gholib, Rinidar Rinidar, Fitriani Fitriani, T. Zahrial Helmi, Sugito Sugito, M. Isa, Nurliana Nurliana, Sri Wahyuni, Dasrul Dasrul and M. Aman Yaman

Veterinary World, 12(7): 1101-1107

ABSTRACT

Background and Aim: Testis and epididymis are male reproductive organs that play an important role in spermatogenesis. These two organs are rich in the content of hormones and other molecules needed in the process of spermatogenesis which affect the quality of the spermatozoa. The objective of this study was to examine the effect of the administration of epididymis and testicular extracts and their combination on testosterone, pituitary adenylate cyclase-activating polypeptide (PACAP), and protamine 1 (PRM1) concentrations in the serum of male chicken.

Materials and Methods: Twenty male chickens (broiler strain Cp707), aged 3 weeks and weighing 800-1000 g, were randomly divided into four different groups including a control group (T0) = injected with 1 ml normal saline and treatment groups: T1 = injected with 1 ml epididymis extract, T2 = injected with 1 ml testicular extract, and T3 = injected with a combination of 1 ml epididymis + 1 ml testicular extract. The experiment was conducted for 13 days and at the end of the study (day 14), the chickens were sacrificed to obtain the serum. Furthermore, the concentrations of testosterone, PACAP, and PRM1 were then measured by using an enzyme-linked immunosorbent assay technique.

Results: The concentrations of PACAP and PRM1 did not show a significant difference between treatment groups (T1, T2, and T3) and control group (T0) (p>0.05). However, the concentration of testosterone showed a significantly higher difference in a group injected with a combination of 1 ml epididymis and 1 ml testicular extracts (T3) compared to the control group (T0) (p<0 .05="" p="">

Conclusion: The administration of epididymis and testicular extracts and their combination did not affect the increase of PACAP and PRM1 concentration. However, a combination of these extracts significantly affects the increase of testosterone concentration in the serum of male chicken.

Keywords: chicken, epididymis and testicular extracts, pituitary adenylate cyclase-activating polypeptide, spermatogenesis, testosterone, protamine 1.