A review: Surveillance of lumpy skin disease (LSD) a growing problem in Asia.

Lumpy skin disease (LSD) is a viral disease caused by lumpy skin disease virus (LSDV), a member of Capripoxvirus, genus of Poxviridae family. It is a transboundary infection of monetary significance that primarily affects water buffaloes and cattle. LSD was known to be once endemic in Saharan regions of Africa but later on reported in central Asian and neighboring countries of Pakistan like, India, Iran and China. It is a vector borne disease and arthropods are believed to be the main perpetrators. It is discernible by its high morbidity and low mortality. Characteristic lumps on skin and high fever are considered as major signs while reduced milk production, infertility, early embryonic death and anorexia are some of the salient clinical manifestations of the disease. Additionally, nodules on mucosa of oro-pharynx, udder, genitalia and rectum are usually observed on examination. This article summarizes LSD outbreaks across Asia during last fifteen years. It is a general consensus amongst the veterinary community that disease is endemic in Pakistan as it shares borders with India, Iran and China where recent outbreaks are reported. Historically Pakistan is free of LSD, however it is at high risk of a LSDV outbreak as neighboring regions are becoming endemic. Vaccination, strict quarantine measures, limited movement of livestock along with vector control could be effective for preventing the spread of the disease. This review aims to summarize the latest developments in the epidemiology of LSD with the focus on transboundary spread, possible emergence and economic implications on Pakistan.

Role of nanotechnology in animal production and veterinary medicine.

Nanotechnology is the discipline and technology of small and specific things that are < 100 nm in size. Because of their extremely miniscule size, any changes in their chemical and physical structure may show higher reactivity and solubility than larger particles. Nanotechnology plays a vital role in every field of life. It is considered one of the most bleeding edge field of scientific research. It has already several applications in a myriad of disciplines while its application in the field of animal production and veterinary medicine is still experimental in nature. But, in recent years, the role of nanotechnology in the aforementioned fields of scientific inquiry has shown great progress. These days, nanotechnology has been employed to revolutionize drug delivery systems and diagnose atypical diseases. Applications of nanoparticle technology in the field of animal reproduction and development of efficacious vaccines have been at the forefront of scientific endeavors. Additionally, their impacts on meat and milk quality are also being judiciously inquired in recent decades. Veterinary nanotechnology has great potential to improve diagnosis and treatment, and provide new tools to this field. This review focuses on some noteworthy applications of nanoparticles in the field of animal production and their future perspectives.

Antibiotic susceptibility profile and synergistic effect of non-steroidal anti-inflammatory drugs on antibacterial activity of resistant antibiotics (Oxytetracycline and Gentamicin) against methicillin resistant Staphylococcus aureus (MRSA).

Non-steroidal anti-inflammatory drugs (NSAIDs) may exhibit antibacterial activity and have synergistic effects with antibiotics. One way to re-sensitize MRSA to resistant antibiotics is by combining with approved non-antibiotics. The study was intended to explore susceptibility of MRSA to various antibiotics and non-antibiotics (NSAIDs) by micro dilution broth method. MRSA isolates were confirmed by PCR (mecA gene) and in-vitro antibiotic susceptibility was determined by disc diffusion tests. Combinations of NSAIDs with resistant antibiotics were also evaluated in in-vivo trial in mice. In-vitro antibiotic susceptibility profile presented 100% resistance to Cefoxitin, 37.5% to Gentamicin and Amikacin, 25% to Oxytetracycline, 12.5% to Tylosin, Fusidic acid and Vancomycin, while Ciprofloxacin, Levofloxacin, Moxifloxacin, Trimethoprim + Sulfamethoxazole and Linezolid were found sensitive. Synergistic effect was observed when resistant antibiotics (Oxytetracycline/Gentamicin) were combined with sensitive antibiotics (Ciprofloxacin/Linezolid). Combination of Non-antibiotics (NSAIDs) with resistant antibiotics revealed that Meloxicam showed partial synergism with both Oxytetracycline and Gentamicin, while Flunixin Meglumine presented synergistic effect with Oxytetracycline and partial synergism with Gentamicin. Diclofenac Sodium revealed additive effect with Oxytetracycline while in case of Gentamicin indifferent effect was observed. In-vivo results showed that combinations of Oxytetracycline with Meloxicam/Flunixin Meglumine and Gentamicin with Meloxicam/Flunixin Meglumine were effective. The study concluded that the resistance against milk borne zoonotic MRSA infections can be successfully addressed by combining resistant antibiotics with NSAIDs. Flunixin Meglumine can be used in combination with oxytetracycline against MRSA infection.

A comprehensive review on past, present and future aspects of canine theileriosis.

Canine theileriosis is a notorious tick borne piroplasmid infection of wild and domestic canines. The causative agent has not yet been accurately classified. PCR studies revealed that causative agent resembles to Theileria genus and thus provisionally named as Theileria annae. The other Theileria species reported in canines is Theileria annulata, Theileria equi and unnamed Theileria specie. This emergent canine infection is considered to be endemic in most of the European countries. However in Asia this disease has not been reported till date. The vectors responsible for transmission of this disease have not been determined. It has been suggested that DNA of Theileria annae has been detected in hard tick Ixodes hexagonus in Northwestern Spain and several other tick species. Clinically canine theileriosis is characterized by severe weakness, fever, hemoglobinuria and anemia. Recently atovaquone or buparvaquone plus azithromycin therapy showed better clinical efficacy. This comprehensive review is intended to summarize the current knowledge on prevalence and epidemiology of canine theileriosis in different countries of the world and associated tick vectors.

The first report on serotyping of bluetongue virus in small ruminants of Khyber Pakhtunkhwa province, Pakistan.

Bluetongue virus (BTV), a member of Orbivirus genus (family Reoviridae), is a non-contagious infection of domestic and wild ruminants. The current study was designed to detect various serotypes of BTV in small ruminants of Khyber Pakhtunkhwa (KPK) province of Pakistan, along with their effects on hemato-biochemical parameters. A total of 408 serum samples in four districts (Mansehra, Abbottabad, Swabi, and Kohat) of KPK from small ruminants were screened based on competitive ELISA (cELISA). A total of 204 (50%) samples were found positive for BTV group-specific antibodies. The seropositive samples were processed for the detection of BTV serotypes through real-time polymerase chain reaction (qPCR). Out of 204 cELISA-positive samples, 60 (29.41%) were found positive through qPCR. Three serotypes [6, 8, 9] were detected from Mansehra District and two from Kohat [2, 8] and Abbottabad [6, 8], while only one from Swabi [8]. The serotype "8" was found consistently in all the four study districts. A significant (p < 0.05) increase in the level of blood urea nitrogen (BUN) and alkaline phosphatase (ALP) was recorded in goats, whereas aspartate aminotransferase (AST) in sheep infected with BTV, compared to healthy animals. The hematological parameters showed significantly (p < 0.05) raised total leucocyte count (TLC) in both sheep and goats, whereas only hematocrit (HCT) value was increased significantly (p < 0.05) in infected sheep. This is the first report on serotyping of BTV among small ruminants in Pakistan.

Equine Granulocytic Anaplasmosis 28 years later.

Equine granulocytic anaplasmosis (EGA) is an important tick borne disease of equines that is caused by Anaplasma phagocytophilum (A. phagocytophilum). The etiological agent has veterinary as well as public health importance because of its zoonotic nature. A. phagocytophilum causes an acute illness in equines with loss of appetite, lethargy, hemorrhages and lameness. Clinically, EGA is diagnosed upon examination of morulae within neutrophils especially granulocytes in the blood. The best diagnostic tool for the detection of EGA is Polymerase chain reaction (PCR). Previous studies suggested that EGA is a self-limiting disease and tetracycline therapy is considered as a best treatment regimen. There is no comprehensive summary on the occurrence and distribution of the infection at global level. Therefore, we intended to provide a comprehensive summary on the prevalence and epidemiology of EGA in different areas of the world. It includes mapping the global distribution of EGA in different areas of the world to identify the endemic regions which may be a source of potential disease outbreak. For this purpose, the published data from 1990 to 2018 on EGA was reviewed and collected by electronic literature search of five databases including Google, Google Scholar, Science Direct, PubMed and Web of Science.

Impact of Maternal Antibodies on Infectious Bronchitis Virus (IBV) Infection in Primary and Secondary Lymphoid Organs of Chickens.

Infectious bronchitis virus (IBV) causes infectious bronchitis disease in chickens. IBV primarily infects the upper respiratory tract and then disseminates to other body systems including gastrointestinal, reproductive, and urinary systems. Unlike original IBV serotypes, the novel IBV variants target lymphoid organs, but information on this is scarce. In this study, we aim to evaluate the impact of the presence of maternal antibodies on IBV infection in primary and secondary lymphoid organs. Maternal antibody free, specific pathogen free (SPF) hens were divided into vaccinated and non-vaccinated groups. The progeny male chicks from these hens were divided into four groups; vaccinated challenged (VC), non-vaccinated challenged (NVC), vaccinated non-challenged (VNC), and non-vaccinated non-challenged (NVNC). The challenge groups were given 1 × 106 embryo infectious dose (EID)50 of IBV Delmarva (DMV)/1639 by the oculo-nasal route and non-challenge groups were given saline. The serum anti-IBV antibody titer was significantly higher in challenged groups compared to non-challenged groups. The IBV genome load was significantly lower in the VC group than NVC group in oropharyngeal and cloacal swabs and in bursa of Fabricius (BF) and cecal tonsils (CT). The histopathological lesion scores were significantly lower in VC group than NVC group in BF and CT. These findings suggest that the presence of maternal antibody in chicks could provide some degree of protection against IBV infection in BF and CT.

Tracking Infection and Genetic Divergence of Methicillin-Resistant Staphylococcus aureus at Pets, Pet Owners, and Environment Interface.

Staphylococcus aureus (S. aureus) has become a leading animal and public health pathogen that keeps on transferring from one host to other, giving rise to newer strains by genetic shifts. The current study was designed to investigate the epidemiology and genetic relatedness of mecA gene in S. aureus isolated from pets, immediate individuals in contact with pets, and veterinary clinic environments. A total of n = 300 samples were collected from different veterinary hospitals in Pakistan using convenience sampling. The collected samples were subjected to microbiological and biochemical examination for the isolation of S. aureus. Methicillin resistance was investigated by both phenotypically using oxacillin disk diffusion assay and by genotypically targeting mecA gene by PCR. PCR amplicons were subjected for sequencing by Sanger method of sequencing, which were subsequently submitted to NCBI GenBank under the accession numbers MT874770, MT874771, and MT874772. Sequence evolutionary analysis and mecA gene characterization was done using various bioinformatics tools. Overall, 33.66% mecA genes harboring S. aureus strains were isolated from all sources (33.33% from pets, 46.0% from surrounding, and 28.0% from immediate contact individuals). The bioinformatics analysis noted that one SNP was identified at position c.253C>A (Transvertion). The phylogenetic tree (two clades) of S. aureus mecA revealed a possibility of inter-transmission of disease between the environment and pets. Frequency of adenine and thymine nucleotide in motifs were found to be the same (0.334). Cytosine and guanine frequency were also the same (0.166). Threonine was replaced by asparagine (p.T84D) in each sample of cat, environment, and human. On the other hand, protein structures ofcat-1 and cat-2 proteins were found identical while cat-3, environmental, and human proteins shared identical structures. The study thus concludes rising circulation of methicillin-resistant S. aureus (MRSA) strains in animal-human-environment interfaces, forecasting the development of novel strains withmodified range of resistance.

MOLECULAR INVESTIGATION AND PHYLOGENETIC ANALYSIS OF ANAPLASMOSIS IN DOGS.

Anaplasmosis is a widespread vector-borne disease affecting dogs, and Anaplasma platys is the major etiological agent of the disease. The study examines anaplasmosis molecular prevalence, related risk factors, and alteration of hematological variables in Anaplasma-affected dogs. A total of 150 blood samples were collected from dogs in the district of Lahore, Pakistan. The samples were screened with PCR targeting the 16S rRNA gene of Anaplasma. Sequencing of samples that were found positive after performing PCR was conducted. A questionnaire was developed to collect epidemiological data on subject dogs, and the information was analyzed with a logistic regression model using SPSS. The current study revealed an 11.34% (17/150) prevalence of anaplasmosis in dogs based on PCR detection. Tick infestation, previous tick history, house hygiene, and tick control status were major risk factors linked with disease occurrence. Red blood cell count, packed cell volume, hemoglobin, and platelet count were decreased significantly (P < 0.05) in Anaplasma-infected dogs. Phylogenetically, the 2 isolates of the current study clustered together, and that cluster was very similar to A. platys isolates from India, Malaysia, and Thailand.

Molecular evidence of Anaplasma infection in naturally affected domestic cats of Pakistan.

Feline anaplasmosis is considered as an emerging tick-borne disease of zoonotic potential. The aim of current study was to investigate the molecular prevalence of anaplasmosis, associated risk factors, and alterations in hematological parameters of domestic cats from Lahore, Pakistan. Blood samples of 100 domestic cats from district Lahore were examined microscopically and the extracted genomic DNA from each sample was processed for the amplification of 16 S rRNA gene of Anaplasma. PCR confirmed isolates were purified for sequencing. The data regarding the risk factors was collected in a predesigned questionnaire and statistically analyzed by logistic regression analysis. The study found a molecular prevalence of 13% (13/100) among analyzed blood samples. The nucleotide analysis of Anaplasmataceae species sequences amplified by PCR showed high resemblance (99%) with isolates from Korea, Japan, Malaysia, Philippines, and India. The potential risk factors found to be significantly associated (p < 0.05) with disease dynamics based on the Chi-Square test were tick infestation on studied animals, previous tick history, tick control status, house hygiene, and housing type. A significant (p < 0.05) decrease in the number of platelets, erythrocytes, hemoglobin level, and pack cell volume was observed in cats suffering from anaplasmosis compared to the healthy ones. The current study is the first report of Anaplasma infection in domestic cats of Pakistan. This study will be effectual in designing the control strategies for this disease.

Molecular Analysis and Risk Factors Associated With Theileria equi Infection in Domestic Donkeys and Mules of Punjab, Pakistan.

Theileriosis is an important disease of economic significance which badly affects the equine husbandry of developing countries. The present study was planned to investigate the molecular prevalence of theileriosis, associated risk factors, and alterations in hematological parameters of donkeys and mules from district Mianwali, Punjab, Pakistan. Blood samples from 150 equids (n = 75 donkeys; n = 75 mules) were examined microscopically, and the genomic DNA from each sample was processed for the amplification of the 18S rRNA gene of Theileria. The polymerase chain reaction confirmed isolates were purified followed by sequencing. The data regarding the analysis of risk factors were collected in a predesigned questionnaire and statistically analyzed by logistic regression analysis. An overall prevalence of 17.33% was noted in this study. Donkeys showed more prevalence followed by mules being 20.0% and 14.7%, respectively. The study isolates showed high resemblance (99%) with isolates from the United States of America, Spain, Brazil, Israel, Cuba, France, South Africa, Korea, Turkey, Tunisia, India, E. Caribbean, and Nigeria. The potential risk factors found to be significantly associated (P < .05) with disease dynamics were tick infestation on study animals, previous tick history, and house hygiene. A significant (P < .05) decrease in the number of platelets, erythrocytes, hemoglobin level, and packed cell volume was observed in donkeys and mules suffering from theileriosis compared with the healthy ones. The study is the first report regarding the molecular characterization of theileriosis in donkeys and mules in Pakistan. The findings will be effectual in designing effective control strategies for this disease in Punjab, Pakistan.

First Report on Molecular Characterization of Anaplasmosis in Small Ruminants in Pakistan.

Anaplasmosis is caused by a Gram-negative obligate intracellular bacterium of the genus Anaplasma with the pathogen having a zoonotic impact. The study aimed to estimate the prevalence of anaplasmosis in Pakistan, to unravel the association of potential risk factors, and to investigate the effect on hematological parameters in affected small ruminants. A total of 150 (n = 75 sheep; n = 75 goats) blood samples were initially screened microscopically and then subjected to PCR targeting the amplification of the 16S rRNA gene fragment of Anaplasma. The PCR-based positive samples were then processed for sequencing. Statistical analysis regarding risk factors was performed using R software. The study revealed an overall 29.33% (44/150) prevalence of anaplasmosis in small ruminants. Sheep had higher (P > 0.05) prevalence (32%) as compared to goats (25.30%). The final statistical model resulting from backward elimination showed only tick infestation as a significant predictor of infection status. The phylogenetic analysis of 16S rRNA gene of Anaplasma spp. revealed 9 study isolates clustered together and showed a close resemblance (99%) with Anaplasma ovis isolate (DQ837600) from Hungary. One of the isolates showed (99%) similarity with the isolate of Anaplasma marginale (MH155594) from Iraq. Furthermore, the hematological parameters pack cell volume, red blood cells, hemoglobin, white blood cells, granulocytes, monocytes, lymphocytes, and platelet count were decreased in Anaplasma-positive animals. This is the first study at the molecular level to characterize Anaplasma spp. in small ruminants of Pakistan, and it will be useful in developing control strategies for anaplasmosis.

Balantidium coli in domestic animals: An emerging protozoan pathogen of zoonotic significance.

Balantidium coli (B. coli) is an emerging ciliated protozoan parasite of zoonotic importance which causes a disease balantidiasis in a variety of host species including pigs, camels, ruminants, equines and even human. This disease has a cosmopolitan distribution with high prevalence rates in tropical and sub-tropical areas of the world due to favorable geo-climatic conditions for the development and survival of the parasite in these regions. Main reservoir hosts for this pathogen are pigs and animals; acquire infection mainly by the ingestion of the food or water contaminated with the porcine feces. The infected animal manifests clinical signs of anorexia, dehydration, profuse watery diarrhea and retarded growth. Wet mount slide prepared from intestinal scrapings and fecal material is used for the identification of trophozoites and cysts stages of this parasite. PCR can also be used to confirm the parasite. Secnidazole, oxytetracycline and metronidazole have varying efficacy against B. coli infection in various domestic animal species. There is no comprehensive literature available on the occurrence and distribution of the infection at international level. Therefore, the published data between 1989 and 2019 regarding this disease is critically analyzed to provide a detailed overview on this pathogen with special emphasis on geographical distribution of B. coli in domestic animals and different therapeutic agents used to treat this infection. This review will pinpoint the endemic regions which may be a source of potential disease outbreaks and will also help in application of more effectual control strategies against balantidiasis.

Sero-epidemiology of bluetongue virus (BTV) infection in sheep and goats of Khyber Pakhtunkhwa province of Pakistan.

Bluetongue virus (BTV) infection is an emerging hazard in small ruminants having socio-economic impacts on animals and associated people. The current study was aimed to estimate the sero-prevalence and associated risk factors in sheep and goat from Khyber Pakhtunkhwa (KP) province of Pakistan. Three distinct zones (northern, central and southern) with four districts (Mansehra, Abbottabad, Swabi, and Kohat) with a higher population of small ruminants were selected. A total of n = 408 sera originating from sheep (n = 212) and goats (n = 196) were randomly collected for detection of BTV group specific antibodies through competitive ELISA (c-ELISA). Univariable and multiple logistic regressions were applied to assess the potential risk factors associated with the occurrence of this disease. Results showed an overall prevalence of 50.00% (CI = 44.17-54.83) of BTV in both sheep and goats with a significant difference (p < 0.05) among different districts. The prevalence of BTV in sheep was found higher (56.60%, CI = 49.6-63.4) than goats (42.86%, CI = 35.8-50.1). The risk factors identified based on chi-square test were; 1-2 year of animals, herd size and location in sheep while, milking status, ticks infestation, location and herd size for goats (p < 0.05). On the basis of univariable analysis, 1-2 year of animals, and location for sheep while, ticks infestation and location for goats (OR > 1). Multiple logistic regressions conferred only herd size and location as potential risk factors (OR > 1) for BTV in sheep and goats. The study concluded higher prevalence of BTV in sheep than the goats, the risk factors were significantly associated with the occurrence of disease, and together ascertaining the needs to design appropriate disease management and control strategies in sheep and goats.