Production and characterization of Newcastle disease antibody as a reagent to develop a rapid immunodiagnostic test tool

Research (Published online: 05-07-2018)

2. Production and characterization of Newcastle disease antibody as a reagent to develop a rapid immunodiagnostic test tool

Dwi Desmiyeni Putri, Ekowati Handharyani, Retno Damajanti Soejoedono, Agus Setiyono and Okti Nadia Poetri

Veterinary World, 11(7): 895-901

ABSTRACT

Aim: This research was conducted to produce and characterize ND antibody as reagent candidate to develop a rapid immunodiagnostic test tool.

Materials and Methods: Four New Zealand White rabbits were used in this study and divided into two groups. First group was injected by Sato ND antigen, and second group was injected by genotype VII ND antigen. This study is divided into three steps: (a) ND antibody production, (b) ND antibody purification, and (c) ND antibody characterization. First group was rabbit injected by Sato NDV (5x108.25 egg lethal doses (ELD)50/ml) and second group was injected by genotype VII NDV (5x106.5 ELD50/ml). Antigen induction was performed by subcutaneous administrated for first (day 1) and second (day 14) injection and intravenous administrated for third (day 30) injection. Blood was collected on day 8 after third injection.

Results: Antibody production increased on second antigen injection and reached a peak on day 9 after second antigen injection. Sato and genotype VII ND antibody can be produced without adjuvant within 38 days with the highest titer 210. Based on antibody titer data, both antigens induced antibody production in a similar trend. The characterization antibody by SDS-PAGE indicated that molecular weight of immunoglobulin G (IgG) is 154.93 kDa (whole IgG), heavy chain 54.39 kDa, and light chain 27.74 kDa. ND antibodies have specificity to homologous and heterologous NDVs in varying virulence.

Conclusion: Sato and genotype VII ND antibodies have been successfully produced within 38 days without adjuvant. Specificity of ND antibodies to NDVs in varying virulence and cross-reaction between Sato ND antibody and genotype VII ND antibody indicates that the characterized ND antibodies can be used as a reagent to develop rapid immunodiagnostic test tools.

Keywords: antibody, cross reaction, reagent, SDS-PAGE.

Characterization of the onset of puberty in Tazegzawt lambs, an endangered Algerian sheep: Body weight, thoracic perimeter, testicular growth, and seminal parameters

Research (Published online: 02-07-2018)

1. Characterization of the onset of puberty in Tazegzawt lambs, an endangered Algerian sheep: Body weight, thoracic perimeter, testicular growth, and seminal parameters

Farid Moulla, Rachid El-Bouyahiaoui, Rebouh Nazih, Norezzine Abdelaziz, Nacera Zerrouki and Mokrane Iguer-Ouada

Veterinary World, 11(7): 889-894

ABSTRACT

Aim: The aim of the present study was to define the onset of puberty in Tazegzawt ram lambs, an Algerian sheep breed in endangered status with a small population in its local area.

Materials and Methods: Body growth (body weight and thoracic perimeter), scrotal circumference (SC), penis development stages, and seminal parameters (volume, concentration, and motility) were measured. Data were recorded at fortnightly intervals in 10 animals from 9 to 49 weeks of age.

Results: On the basis of seminal analyses, puberty occurred between 29 and 45 weeks of age. At 29 weeks of age, 30% of lambs reached puberty, and at 45 weeks of age, puberty was observed in 100% of the analyzed animals. Body weight appeared as the most determinant factor, and the onset of puberty was observed when animals reached 43.2±6.4 kg body weight with 25.8±3.7 cm of SC. Seminal analyses revealed that all parameters increased regularly from puberty onset except for sperm concentration. The mean semen volume during the study period was 0.48±0.33 mL with 0.84±0.6 mL at 37 weeks of age. Sperm concentration evolved similarly as semen volume; at 29 and 43 weeks of age, the sperm concentration was 942x106 and 1904x106 spermatozoa/mL, respectively. Kinematic parameters including the percentage of motility, the percentage of progressive motility, and gametes velocities as determined by Computer-Aided Sperm Analyzer showed the highest values at 49 weeks of age.

Conclusion: The current results revealed that, in Tazegzawt ram lambs, puberty occurs between 29 and 45 weeks when animals reach 43.2±4.6 kg body weight.

Keywords: body weight, puberty onset, scrotal circumference, seminal parameters, Tazegzawt breed sheep.

Reproduction performance and blood biochemical parameters in dairy cows: Relationship with oxidative stress status

Research (Published online: 30-06-2018)

23. Reproduction performance and blood biochemical parameters in dairy cows: Relationship with oxidative stress status

Sofiane Boudjellaba, Lynda Ainouz, Safia Tennah, Soraya Temim and Mokrane Iguer-Ouada

Veterinary World, 11(6): 883-888

ABSTRACT

Background and Aim: During the last decades, reproduction performances declined dramatically worldwide, but little is known concerning the involvement of oxidative stress as a causative factor. Oxidative stress may act at different levels, with negative impacts on cell membrane integrity and other active molecules with potential subsequent effects on reproduction. The aim of the current study was to investigate the oxidative stress status in cows according to their reproductive performances.

Materials and Methods: Peripheral blood concentration of two oxidative stress biomarkers, glutathione S-transferase (GST) and malondialdehyde (MDA), and other biochemical parameters (glucose, total lipids, cholesterol, triglycerides, albumin, total proteins, calcium, urea, creatinine, direct bilirubin, alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase) were determined in 40 healthy cows. Body condition score (BCS), calving to first service interval (FSI), calving to conception interval (CCI), and the number of service per conception (SPC) were simultaneously recorded for each cow.

Results: Concerning FSI, three groups were established: Group 1 (from 44 to 60 days), Group 2 (from 60 to 70 days), and Group 3 (from 70 to 80 days). For CCI, two groups were considered: Group 1 (<110 days) and Group 2 (>110 days). MDA showed significant high values only in cows with the lowest BCS (1.5) compared to cows with BCS note of 2.5 and 3.5. No significant difference was observed in cows oxidative stress status (MDA and GST) according to reproductive performances (FSI, CCI, and SPC) in all studied groups.

Conclusion: The results revealed relatively altered oxidative stress status in cows with abnormal reproductive performances; however, no significant difference was recorded whatever the considered reproductive parameter.

Keywords: cow, glutathione S-transferase, malondialdehyde, reproduction performance.

Hematological parameters and selected intestinal microbiota populations in the Indonesian indigenous crossbred chickens fed basal diet supplemented with multi-strain probiotic preparation in combination with vitamins and minerals

Research (Published online: 29-06-2018)

22. Hematological parameters and selected intestinal microbiota populations in the Indonesian indigenous crossbred chickens fed basal diet supplemented with multi-strain probiotic preparation in combination with vitamins and minerals

Sugiharto Sugiharto, Turrini Yudiarti, Isroli Isroli, Endang Widiastuti and Hanny I. Wahyuni

Veterinary World, 11(6): 874-882

ABSTRACT

Aim: The aim of this study was to investigate the effect of dietary supplementation with multi-strain probiotic preparation in combination with vitamins and minerals on the hematological parameters and selected intestinal microbiota populations in the Indonesian indigenous crossbred chickens.

Materials and Methods: A total of 240 one-day-old Indonesian indigenous crossbred chicks were raised for 10 weeks. The chicks were distributed to one of four groups, i.e., chicks receiving basal diet without any additive (CONT), chicks receiving basal diet with 0.04% of zinc bacitracin (AGP), chicks receiving basal diet with 0.01% of commercial probiotic Bacillus subtilis preparation (PROB1), and chicks receiving basal diet with 0.5% of multi-strain probiotic preparation in combination with vitamins and minerals (PROB2). Blood was collected on the week 8, while the internal organs and eviscerated carcasses were collected on the week 10.

Results: PROB2 tended (p=0.09) to have a lower body weight (BW) compared to CONT chicks. Feed conversion ratio was higher (p<0.05) in PROB1 and PROB2 compared to CONT birds. The number of thrombocytes tended (p=0.09) to be higher in CONT than in other groups. Antibody titer against Newcastle disease virus vaccine was higher (p<0.05) in PROB1 and PROB2 than in CONT group. Serum triglyceride concentration was lower (p<0.05) in PROB2 than in other birds. AGP chicks had lower (p<0.05) serum total protein and globulin concentrations than CONT and PROB1 chicks. Serum albumin level was lower (p<0.05) in PROB2 than in CONT and PROB1 birds. Albumin to globulin ratio tended (p=0.06) to be higher in AGP than in other birds. Lactose-negative Enterobacteriaceae tended (p=0.07) to be lower in PROB1 and PROB2 than in CONT group. PROB1 and PROB2 tended (p=0.06) to have greater lactic acid bacteria (LAB) population than in CONT and AGP birds.

Conclusion: Multi-strain probiotic preparation in combination with vitamins and minerals was able to improve immune response and control the potentially pathogenic bacteria. However, the additive could not improve the growth performance of the Indonesian indigenous crossbred chickens.

Keywords: growth performance, Indonesian indigenous crossbred chickens, intestinal bacteria, minerals, multi-strain probiotic, vitamins.

Extended-spectrum β-lactamases producing multidrug resistance Escherichia coli, Salmonella and Klebsiella pneumoniae in pig population of Assam and Meghalaya, India

Research (Published online: 29-06-2018)

21. Extended-spectrum β-lactamases producing multidrug resistance Escherichia coli, Salmonella and Klebsiella pneumoniae in pig population of Assam and Meghalaya, India

A. Lalruatdiki, T. K. Dutta, P. Roychoudhury and P. K. Subudhi

Veterinary World, 11(6): 868-873

ABSTRACT

Aim: The present study was conducted to record the prevalence of extended spectrum β-lactamases (ESBLs) producing Escherichia coli, Salmonella spp., and Klebsiella pneumoniae from pig population of Assam and Meghalaya and to record the ability of the resistant bacteria to transfer the resistance genes horizontally.

Materials and Methods: Fecal samples (n=228), collected from pigs of Assam (n=99) and Meghalaya (n=129), were processed for isolation and identification of E. coli and Salmonella spp. All the isolates were tested for ESBLs production by double disc synergy test (DDST) followed by screening for ESBLs producing genes (blaTEM, blaSHV, blaCTX-M, and blaCMY) by polymerase chain reaction (PCR). Possible transfer of resistance encoding genes between enteric bacterial species was carried out by in vitro and in vivo horizontal gene transfer (HGT) method.

Results: A total of 897 enteric bacteria (867 E. coli and 30 Salmonella) were isolated and identified. Altogether 25.41% isolates were confirmed as ESBL producers by DDST method. Majority of the isolates were E. colifollowed by Salmonella. By PCR, 9.03% isolates were found positive for at least one of the target resistance genes. blaSHV was absent in all the isolates. blaCMY was the most prevalent gene. All the E. coli isolates from Assam were negative for blaTEM. A total of 2.76% isolates were positive for blaTEM + blaCMY. On the other hand, 0.67% isolates were positive for blaCTX-M + blaCMY genes. Only 0.33% isolates carried all the three genes. Altogether, 4.68% bacteria carried the resistance encoding genes in their plasmids. blaTEM gene could be successfully transferred from Salmonella (donor) to E. coli (recipient) by in vitro (5.5-5.7x10-5) and in vivo (6.5x10-5to 8.8x10-4) methods. In vivo method was more effective than in vitro in the transfer of resistance genes.

Conclusion: The pig population of Assam and Meghalaya are carrying multidrug resistance and ESBLs producing E. coli and Salmonella. The isolates are also capable to transfer their resistance trait to other bacterial species by HGT. The present finding could be considered as a serious public health concern as similar trait can also be transmitted to the human commensal bacteria as well as pathogens.

Keywords: Enterobacteriaceae, multidrug resistance, North East India, pigs.

Molecular characterization and antibiotic resistance patterns of avian fecal Escherichia coli from turkeys, geese, and ducks

Research (Published online: 27-06-2018)

20. Molecular characterization and antibiotic resistance patterns of avian fecal Escherichia coli from turkeys, geese, and ducks

Nokukhanya Dube and Joshua Mbanga

Veterinary World, 11(6): 859-867

ABSTRACT

Background and Aim: Avian fecal Escherichia coli (AFEC) are considered to be the natural reservoir of pathogenic strains in extraintestinal infections as such characterization of AFEC gives insight into the spread of the potential pathogenic lineage. The aim of the study was to investigate the reservoirs of avian pathogenic E. coli (APEC) from fecal samples of healthy ducks, geese, and turkeys by determining the antibiotic resistance patterns of AFEC isolates from turkeys, geese and ducks and characterization of the isolates using virulence genes, plasmid profiles, and phylogenetic grouping.

Materials and Methods: The disc diffusion method was used to determine antibiotic resistance of 100 AFEC isolates from turkeys (9), geese (29), and ducks (62) to 8 antibiotics. Molecular characterization of the isolates was done by multiplex polymerase chain reaction to investigate the presence of 12 virulence genes, plasmid profiling, and phylogenetic grouping based on the 16S rRNA sequences.

Results: Antibiogram profiles indicated maximum resistance to cloxacillin (100%) and bacitracin (100%) for all AFEC isolates and high sensitivity to ciprofloxacin; however, all isolates exhibited multi-drug resistance. The AFEC isolates from turkeys (6) and geese (12) did not contain virulence genes. The frz (3.7%), sitD (29.6%), and fimH (92.5%) were detected in the duck isolates. None of the isolates had the KpsM, iutA, vat, sitA, hlyF, pstB, ompT, uvrY, and sopB genes. Plasmid profiling gave four plasmid profiles with the plasmids ranging from 1.5 to 55 kb. Phylogenetic analysis of 16S rRNA sequences revealed similarities between AFEC isolates from the different poultry species, as the isolates did not cluster according to avian species.

Conclusion: AFEC isolates are potential reservoirs of APEC as they contain some of the virulence genes associated with APEC. Multidrug resistance is high in AFEC isolated from healthy birds. This is a public health concern.

Keywords: antibiotic resistance, avian fecal Escherichia coli, poultry, virulence gene.

Insights into pH regulatory mechanisms in mediating spermatozoa functions

Review (Published online: 26-06-2018)

19. Insights into pH regulatory mechanisms in mediating spermatozoa functions

Abhishek Kumar Mishra, Akshay Kumar, Dilip Kumar Swain, Sarvajeet Yadav and Rajesh Nigam

Veterinary World, 11(6): 852-858

ABSTRACT

Regulation of pH in spermatozoa is a complex and dynamic process as sperm cells encounter different pH gradients during their journey from testes to the site of fertilization in female genital tract. The precise regulations of pH in sperm cells regulate the sperm functions such as motility, hyperactivity, capacitation, and acrosome reaction. Electrophysiological, pharmacological, and molecular studies have revealed the presence of different ion channels and exchanger systems which regulate intracellular pH in sperm cells as well as regulate sperm functions. Recent studies also have shown the potential involvement of pH in the regulation of fertility competence of sperm cells, and alterations in pH have shown to impede sperm functions. This mini-review discusses the probable mechanisms involved in pH regulation in sperm cells and how pH is involved in regulation of various sperm functions.

Keywords: acrosome reaction, capacitation, fertilization, pH, sperm motility, spermatozoa.