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.

Preliminary study on the tick population of Benin wildlife at the moment of its invasion by the Rhipicephalus microplus tick (Canestrini, 1888)

Research (Published online: 25-06-2018)

18. Preliminary study on the tick population of Benin wildlife at the moment of its invasion by the Rhipicephalus microplus tick (Canestrini, 1888)

Kossi Justin Adinci, Yao Akpo, Camus Adoligbe, Safiou Bienvenu Adehan, Roland Eric Yessinou, Akoeugnigan Idelphonse Sode, Guy Appolinaire Mensah, Abdou Karim Issaka Youssao, Brice Sinsin and Souaibou Farougou

Veterinary World, 11(6): 845-851

ABSTRACT

Background and Aim: Rhipicephalus microplus (Rm) is one of the most problematic livestock tick species in the world. Its rapid propagation and resistance to acaricides make it control difficult in the sub-region and Benin particularly. The aim of this work was to check its presence in wildlife and to confirm the possible role of reservoir wildlife may play in the propagation of the parasite. This will help to design more efficient control strategy.

Materials and Methods: This study was conducted from February to March 2017 in the National Parks of Benin (Pendjari and W Park) and wildfowl's assembly and selling point in Benin. Ticks were manually picked with forceps from each animal after slaughtering by hunters then stored in 70° ethanol. Collected ticks were counted and identified in the laboratory using the identification key as described by Walker.

Results: Overall, seven species of ticks (Amblyomma variegatum, Boophilus decoloratus, Rm, Boophilus spp., Hyalomma spp., Rhipicephalus sanguineus, Rhipicephalus spp.) were identified on nine wild animal species sampled (Cane rat, wildcat, Hare, Doe, Cricetoma, Buffalo, Buffon Cobe, and Bushbuck and Warthog). The average number of ticks varies from 3 to 6 between animal species, 3 to 7 between localities visited, and 2 to 5 between tick species. However, these differences are statistically significant only for localities. Considering tick species and animal species, the parasite load of Rm and Rhipicephalus spp. is higher; the buffalo being more infested. The analysis of deviance reveals that the abundance of ticks observed depends only on the observed localities (p>0.05). However, the interactions between animal species and localities on the one hand and between animal and tick species on the other hand, although not significant, have influenced the abundance of ticks as they reduce the residual deviance after their inclusion in the model.

Conclusion: This study reported the presence of Rm in wildlife of Benin and confirmed its role in the maintenance and spread of the parasites. It is, therefore, an important risk factor that we must not neglect in the epidemiological surveillance and ticks control strategies in the West African sub-region and particularly in Benin.

Keywords: Benin, Rhipicephalus microplus, ticks, wild animals.

Brucella seropositivity in chicken and risk factors for Brucella infection at the animal-human interface in Anambra State, Nigeria

Research (Published online: 24-06-2018)

5. Brucella seropositivity in chicken and risk factors for Brucella infection at the animal-human interface in Anambra State, Nigeria

Joseph Ikechukwu Onunkwo, Emmanuel Okechukwu Njoga, Ugochinyere Juliet Njoga, Emmanuel Ezeokafor and Samuel Okezie Ekere

International Journal of One Health, 4: 28-34

ABSTRACT

Aim: Brucellosis is an important bacterial zoonosis devastating both animal and human populations in many parts of the world. A seroepidemiological study of avian Brucella infection was conducted to determine the disease prevalence, risk factors, and hence the role of chicken in the epidemiology of brucellosis in Anambra State, Nigeria.

Materials and Methods: Rose Bengal plate test was used to test for Brucella antibody in sera samples collected from 410 chickens surveyed. The interview schedule was used to elicit information on the socioeconomic status, awareness of brucellosis and predisposing practices of poultry farmers, live bird sellers, and poultry carcass processors in the study area.

Results: An overall seroprevalence of 3% was recorded. Sex (female), free-range management system, breed (indigenous breed), and mix farming were the determinants of avian brucellosis in the state. Risk factors that may enhance human Brucella infection at the animal-human interface are non-use of personal protective clothing; poor awareness on brucellosis and methods of the disease spread or control, cohabitation with animals, and eating while on duty.

Conclusion: Chicken may be among the reservoirs of Brucella infection in Anambra State. There is an urgent need for an effective control program against brucellosis in the study area, using a coordinated One Health approach bearing in mind the public health and economic consequences of brucellosis.

Keywords: Brucella antibodies, Brucella infection, Brucella, chickens, humans, risk factors.

Leptospirosis seropositivity and its serovars among cattle in Northeastern Malaysia

Research (Published online: 23-06-2018)

17. Leptospirosis seropositivity and its serovars among cattle in Northeastern Malaysia

Aziah Daud, Nik Mohd Hafiz Mohd Fuzi, Mohd Mokhtar Arshad, Suratan Kamarudin, Wan Mohd Zahiruddin Wan Mohammad, Fairuz Amran and Nabilah Ismail

Veterinary World, 11(6): 840-844

ABSTRACT

Background: Leptospirosis is a zoonotic disease that infects human and livestock which causes economic losses to the farmers. It has been reported as one of the causes of reproductive failure in cattle and other ruminants, determining abortions, stillbirth, weak newborns, and decrease in their growth rate and milk production.

Aim: The objectives of this study were to determine the leptospirosis seroprevalence and to identify the predominant infecting serovars among cattle.

Materials and Methods: A cross-sectional study involving 420 cattle from six randomly selected districts in Kelantan was conducted. A serological test using the microscopic agglutination test was conducted in the Institute of Medical Research with a cutoff titer for seropositivity of ≥1:100.

Results: The overall prevalence of leptospirosis seropositivity among cattle in this study was 81.7% (95% confidence interval: 63.5, 80.1). The most common reaction obtained with the sera tested was from the serovar Sarawak with 78.8%.

Conclusion: A high seroprevalence of leptospiral antibodies was found among cattle in Northeastern Malaysia. These findings urge that more studies are required to determine the reasons for the high seroprevalence among the cattle along with its transmission and pathogenicity of the local serovar Sarawak.

Keywords: cattle, leptospirosis, microscopic agglutination test, serovar Sarawak.

Effects of meniran (Phyllanthus niruri L.) administration on leukocyte profile of broiler chickens infected with Mycoplasma gallisepticum

Research (Published online: 22-06-2018)

16. Effects of meniran (Phyllanthus niruri L.) administration on leukocyte profile of broiler chickens infected with Mycoplasma gallisepticum

Sri Hidanah, Emy Koestanti Sabdoningrum, Retno Sri Wahjuni and Sri Chusniati

Veterinary World, 11(6): 834-839

ABSTRACT

Aim: This study aimed to evaluate the effects of Meniran extract (Phyllanthus niruri L.) administration on leukocyte profile of broiler chickens infected with Mycoplasma gallisepticum.

Materials and Methods: Thirty broiler chickens, 21 days old were divided into five treatment groups. P0 (-): Chickens without any treatment; P0 (+), P1, P2, and P3: Chickens were infected with M. gallisepticum 108cells/ml/animal orally, then given no treatment, Meniran extract 60%, 62.5%, and 65% orally at a dose of 1 ml/kg body weight, respectively. The treatment of Meniran extract was given for 7 days.

Results: Leukocyte count with the lowest number showed in Group P0 (-) and Group P3 (p>0.05). Increased number of basophils was found in Group P0 (+), Group P1, and Group P2. The highest number of heterophils was found in Group P0 (+) and was significantly different from Group P0 to P3 (p<0.05). The same pattern was also seen in the number of lymphocytes in all treatment groups. The number of monocytes showed no significant difference between all treatment groups (p>0.05).

Discussion: Increased the number of leukocytes is often observed in inflammation due to general infections, trauma, or toxicity. Shifting in the number of heterophile or lymphocytes, an increase in the number of monocytes, basophils, and eosinophils may also be associated with various infectious or inflammatory conditions. Heterophils play a role as an antibacterial defense through several effective mechanisms. When infections and inflammation occur, the heterophils will increase to phagocytosis microbe.

Conclusion: It can be concluded that Meniran extract (P. niruri L.) at a dose of 65% can decrease the total number of leukocytes in broilers infected with M. gallisepticum.

Keywords: chicken, leukocyte, Mycoplasma gallisepticum, Phyllanthus niruri L.

Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism

Research (Published online: 22-06-2018)

15. Species adulteration in raw milk samples using polymerase chain reaction-restriction fragment length polymorphism

Rania M. Ewida and Doaa Safwat M. Abd El-Magiud

Veterinary World, 11(6): 830-833

ABSTRACT

Background and Aim: Milk adulteration is pivotal because it leads to worse effects in public health as human adverse reactions with clinical signs ranged from gastrointestinal signs to anaphylactic shock. This study was carried out to estimate the prevalence of adulteration in buffalo's milk sold in Assiut City, Egypt.

Materials and Methods: A total of 50 raw buffalo's milk samples were collected and examined for adulteration by addition of cow's milk. The examination carried out by applying polymerase chain reaction-restriction fragment length polymorphism technique using cytochrome b (cyt b) gene primers and Hinf I enzymes. The size of target gene was 360 bp in both animal species and amplicon can be digested using Hinf I enzyme, this restriction enzyme divided the essential band to clear three bands at 360, 210, and 150 bp in cows' milk, while, the enzyme could not be cleaved the amplicon in buffalo's samples.

Results: The obtained results cleared that the incidence of adulteration of buffalo's milk very high percentage reaches 90%.

Conclusion: It could be concluded that the raw buffalo's milk sold in Assiut City subject to fraudulent practice and thus can lead to public health hazards.

Keywords: buffalo's milk, cyt b gene, Hinf I enzyme, medicolegal, milk adulteration, polymerase chain reaction-restriction fragment length polymorphism.

Antiviral activity of Acanthaster planci phospholipase A2 against human immunodeficiency virus

Research (Published online: 20-06-2018)

14. Antiviral activity of Acanthaster planci phospholipase A2 against human immunodeficiency virus

Anondho Wijanarko, Kenny Lischer, Heri Hermansyah, Diah Kartika Pratami and Muhamad Sahlan

Veterinary World, 11(6): 824-829

ABSTRACT

Aim: Investigation of antiviral activity of Acanthaster planci phospholipase A2 (AP-PLA2) from moluccas to human immunodeficiency virus (HIV).

Materials and Methods: Crude venom (CV) and F20 (PLA2 with 20% fractioned by ammonium sulfate) as a sample of PLA 2 obtained from A. planci's extract were used. Enzymatic activity of PLA2 was determined using the degradation of phosphatidylcholine (PC). Activity test was performed using in vitro method using coculture of phytohemagglutinin-stimulated peripheral blood mononuclear cell (PBMC) from a blood donor and PBMC from HIV patient. Toxicity test of AP-PLA2 was done using lethal concentration required to kill 50% of the population (LC50).

Results: AP-PLA2 F20 had activity and purity by 15.66 times bigger than CV. The test showed that the LC50 of AP-PLA2 is 1.638 mg/ml. Antiviral analysis of AP-PLA2 in vitro showed the inhibition of HIV infection to PBMC. HIV culture with AP-PLA2 and without AP-PLA2 has shown the number of infected PBMC (0.299±0.212% and 9.718±0.802%). Subsequently, RNA amplification of HIV using reverse transcriptase-polymerase chain reaction resulted in the decrease of band intensity in gag gene of HIV.

Conclusion: This research suggests that AP-PLA2 has the potential to develop as an antiviral agent because in vitro experiment showed its ability to decrease HIV infection in PBMC and the number of HIV ribonucleic acid in culture.

Keywords: Acanthaster planci, antiviral activity, human immunodeficiency virus, Indonesia, phospholipase.

Humoral and cellular immune response induced by antigenic protein of Sarcoptes scabiei var. caprae

Research (Published online: 19-06-2018)

13. Humoral and cellular immune response induced by antigenic protein of Sarcoptes scabiei var. caprae

Nunuk Dyah Retno Lastuti, Wiwik Misaco Yuniarti, Poedji Hastutiek, Lucia Tri Suwanti and Dony Chrismanto

Veterinary World, 11(6): 819-823

ABSTRACT

Aim: Scabies is one of the most important diseases in goats and caused by a complex hypersensitivity process that involves both humoral and cell-mediated immune responses. This phenomenon shows that the variety of Sarcoptes scabiei has different characteristics of specific antigenic protein or different immune-dominant. This research aims to detect the humoral and cellular immune response of rabbits which were immunized with the protein of S. scabiei var. caprae.

Materials and Methods: This research was done as follows, identification and collection of Sarcoptes scabiei var caprae from goat infected with scabies, separation of protein antigen from S. scabiei mites with ultrasonic sonicator, measurement of protein content with spectrophotometry, rabbit injection with 500 μg dose of antigen protein which was repeated 5 times (5x booster) every 2 weeks. Measurement of IgG titer using indirect ELISA, whereas to detect the expression of cellular immune response (TLR-9, CD4, and CD8) using Direct Immunofluorescence assay.

Results: Based on the statistical analysis, it showed that there was a significant enhancement (p<0.05) for optical density value or antibody titer and cellular immune response was shown by TLR-9, CD4, and CD8 expression in rabbit T lymphocytes which appear yellow to green fluorescent color using fluorescence microscope. The amount of fluorescence T lymphocytes showed a significant difference (p<0.05) between the control and various boosters.

Conclusion: Antigenic protein of S. scabiei var. caprae contains ligands, which are involved in the pathogen-associated molecular pattern that has an ability to induce humoral and cellular immune response in rabbit. Specifically, that TLR-9 is not only involved in innate immunity but also in adaptive immunity and can be used as alternative adjuvant development research.

Keywords: antigenic protein, CD4, CD8, immunoglobulin G, Sarcoptes scabiei var. caprae, toll-like receptor-9.