Brucellosis is a bacterial disease due to Brucella melitensis, considered a zoonotic agent affecting humans and animals, especially in areas with high disease occurrence, south Khyber Pakhtunkhwa, Pakistan. This work was designed to evaluate the molecular rate of B. melitensis in humans and cattle species and also to perform a phylogenetic analysis between both species. A cross-sectional survey involving 800 participants, including 600 cattle and 200 human participants, underwent blood sample collection with conventional PCR and IS711 locus PCR amplification and Sanger sequencing. The findings detected Brucella melitensis in 37 of the 800 samples, with a molecular prevalence of 3.1 % in cattle and 9 % in humans. The molecular trees play a role in zoonotic transmission and point to the necessity of a further unified approach toward the management of brucellosis in both humans and animals. This is further backed by the use of 95 % C.I for the prevalence rates making the results statistically robust. This research shows that using the IS711 insertion sequence is an efficient and selective method for identifying Brucella species.

There is so far no available data about how the additive, synergistic, or antagonistic effects of the combined form of alpha-lactalbumin (α-La) and bacteriophages might modulate the cellular milieu of the host-pathogen interface. A co-culture of colonocytes and hepatocytes was stimulated with Pseudomonas aeruginosa PAO1 in the presence of KPP22 phage and incubated for 6 hours in medium alone or medium supplemented with bovine milk-origin α-La. The combination of KPP22 phage and α-La significantly inhibited P.a PAO1-elicited secretion of IL-1β, IL-6, and ICAM-1, which are the mediators and enzymes associated with the inflammatory response to an infectious-inflamed milieu. Cell viability was higher in the P.a PAO1+ KPP22 phage group compared to the P.a PAO1alone group. KPP22 phage and α-La, either alone or in combination, rescued P.a PAO1-induced aberrant PGE1/PGE2 production ratios. The convergence of ingested α-La and phages mitigates pro-inflammatory mediators. α-La leads to an increased sensitivity of opportunistic pathogenic bacteria to phages. Structural, functional, or immunological similarities between ingested α-La and phages play an important role in the mitigation of infection-driven pathobiological processes.

This study aimed to investigate the concurrent infection of Pasteurella multocida (P. multocida) type B:2, which causes Hemorrhagic Septicemia (HS), with cases of Foot and Mouth Disease (FMD) outbreaks in cattle in Bangladesh between March and December 2023. Samples were collected from 11 distinct outbreak areas, totaling 102 samples. These included 54 FMD samples (saliva, tissue epithelium, and morbid tissues such as lung, spleen, and heart) and 54 HS samples (nasal swabs and morbid tissues) from 50 cattle of various ages and sexes, all showing clinical signs of suspected concurrent HS and FMD infection. After sample processing, molecular detection of FMDV and its serotypes was performed using Reverse Transcriptase PCR (RT-PCR) with universal and serotype-specific primers. The HS-causing agent, P. multocida type B:2, was initially identified through cultural and morphological characteristics on various media, followed by Gram's and methylene blue staining, biochemical tests, and pathogenicity tests through inoculation of isolates into mice. Finally, molecular detection of P. multocida type B:2 was confirmed using PCR with specific primers. Forty-five (83 %) of the 54 FMD suspected samples tested positive for FMDV, with 53 % of these positive for serotype 'O,' 17 % for serotype 'A,' and 6 % for mixed serotypes 'O' and 'A.' Among the FMDV-positive samples, 17 (38 %) of the HS-suspected samples tested positive for concurrent infection with P. multocida type B:2. The study reveals that FMDV-induced acute immunosuppression in cattle can lead to complications from concurrent infections, particularly those caused by P. multocida type B:2, resulting in HS alongside FMD.

Feline panleukopenia (FPL) in cats is caused by either feline parvovirus (FPV) or canine parvovirus (CPV-2), which belong to the same species "Protoparvovirus carnivoran 1". While FPV is widely recognized as the principal cause of FPL, CPV-2 has been detected at a higher rate than FPV in sick cats in a recent Egyptian study. To assess this conflict, the present study aimed to determine which Protoparvovirus carnivoran 1 is commonly associated with FPL in Egyptian cats. From Dec-2022 to Jan-2024, 43 cats presenting with acute gastroenteritis and testing positive for FPL using in-clinic assay, SNAP® parvo, were tested for Protoparvovirus carnivoran 1 DNA using conventional PCR. Typing of Protoparvovirus carnivoran 1 was conducted by partial VP2 gene sequencing. Additional epidemiological aspects of the disease were investigated, including seasonal pattern, case-fatality rate, median survival time to death, and the association between FPL outcomes and selected factors like age, sex, vaccination status, and clinical signs (vomiting and diarrhea). All cats tested positive for Protoparvovirus carnivoran 1 DNA and FPV was detected in all cats with strong PCR amplicons (n=39). The following seasonal pattern was recorded: cases emerging in autumn, peaking during winter, declining in spring, and disappearing in summer. The case-fatality rate was 41.6 %, and the median time to death was two days. None of the studied factors affected FPL outcomes. In conclusion, FPL in Egyptian cat populations is primarily caused by FPV, not CPV-2, and is particularly prevalent in winter.

Anaplasma marginale, a tick-borne obligate intracellular rickettsia is incriminated to cause heavy economic losses throughout the tropical and subtropical regions, including India. However, studies highlighting the phylogeography and demographic dynamics of A. marginale are very scant from India. Thus, the present study assessed the cladistics and population structure of Anaplasma marginale based on the genes encoding the major surface proteins (MSP) 4 and 5. The cladistics based on phylogenetic tree including the sequences generated herein with the GenBank archived sequences of A. marginale was performed. The relationship between A. marginale haplotypes based on both the genetic markers was estimated by performing median joining (MJ) haplotype network analysis. Demographic dynamics involving population diversity indices and neutrality indices was also performed. The cladistics and MJ haplotype network analysis corresponding to both the markers demonstrated the presence of three distinct clades (1-3) of A. marginale. Population structure analysis revealed low nucleotide (0.00236 ± 0.00064 and 0.00955 ± 0.00101) and haplotype (0.321 ± 0.073 and 0.493 ± 0.083) diversities for the MSP4 and MSP5 genes, respectively. High genetic structuring and low gene flow [Nm values ranging between 0.044481 and 0.208337 for the MSP4 gene and 0.032735 (clades 1 and 2) for the MSP5 gene] was also recorded among the different clades of A. marginale, based on both genetic markers. The present study highlighted the presence of different clades of A. marginale distributed worldwide. The isolates circulating in north India belonged to the dominant clade prevalent throughout the globe.

A zoonotic disease unique to South Asia, brucellosis causes major public health and financial problems, notably in Pakistan. This research aimed to ascertain the molecular frequency of Brucella abortus and Brucella melitensis in humans and cattle and to pinpoint related risk factors in the Southern Khyber Pakhtunkhwa, Pakistan, districts. A total of 800 blood samples, 600 from cattle and 200 from humans, were gathered and examined using traditional PCR targeting the IS711 locus. In humans, the molecular frequency of Brucella melitensis and Brucella abortus was 16.5 %; in cattle, it was 9.1 %. Risk factor analysis in cattle revealed that seasonality, geographic location, sex, and insemination method significantly impacted brucellosis prevalence· Higher risk was observed in female cattle, those naturally inseminated, and during the summer months, particularly in regions such as Bannu· In humans, univariate logistic regression identified potential risk factors, including age (25-50 years), rural residency, lower socioeconomic status, female gender, direct contact with animals, and consumption of raw animal products. However, none of these factors reached statistical significance. The findings underscore the importance of continuous surveillance and targeted preventive measures to control brucellosis transmission between cattle and humans. Understanding the molecular prevalence and risk factors can guide public health strategies and improve livestock management practices to mitigate this zoonotic disease.

Avian Polyomaviruses are imposing severe health problems in budgerigars, non-budgerigar Psittacine species, and non-psittacine species all over the world, including Pakistan. It marks future challenges for aviculturists and pet store owners, causing significant financial losses. This study emphasizes the occurrence and molecular characterization of polyomaviruses in budgerigars and non-budgerigar Psittacine species. Thirty-five feather Samples of adult birds and 15 tissue samples of deceased birds were collected for the detection of Avian polyomavirus based on the VP1 gene. Screening of samples by PCR revealed the presence of 550 bp VP1 gene in deceased nestlings of two lovebirds and four budgerigars, while the feather samples of adult birds were all negative for VP1 gene. The overall positive rate of APV in Psittacine birds was 6/50 (12 %), and the distribution frequency of virus among species was 4/19 (20 %) in Budgerigars and 2/31 (6.4 %) in non-budgerigar. Positive samples were subjected to partial sequencing which showed a nucleotide similarity index of VP1 gene between 97.46 % & 99.6 % with reference sequences in GenBank. The main problem that researchers are dealing with is the scarcity of data on the prevalence and identification of APV in Pakistan. This study is a milestone for further research on APV for the diagnosis and development of vaccines.

The Crimean-Congo haemorrhagic fever (CCHF) is a tropical viral zoonosis first reported in Spain with 17 detected cases since 2010-2024. Health professionals, including veterinarians, play a crucial role in controlling and preventing this disease. This study aimed to analyse and compare the knowledge, perceptions, and attitudes of veterinary students and professionals in Spain regarding CCHF and zoonoses in general. Additionally, the study highlighted the value of epidemiological surveys as a tool for identifying knowledge gaps related to specific diseases. A cross-sectional survey was conducted from January to April 2024 among 4th and 5th-year veterinary students, Masters and Doctorate students, and veterinarians from the public and private sectors using a validated online questionnaire. A total of 135 individuals participated, 80.7 % professionals and 19.3 % students. The study found that 76.9 % of students and 64.2 % of veterinarians had good or very good knowledge of CCHF, yet significant gaps remained, particularly in epidemiology and diagnosis. Notably, only 34.8 % had good or very good knowledge of the disease's situation in Spain, and many respondents felt their training and the information provided by official sources were inadequate. Additionally, the perception of infection risk from pets and attitudes towards preventing zoonoses through medication, vaccination, or regular analysis were concerning. Despite CCHF being an emerging disease in Spain, this survey-the first in Spain and Europe-reveals that veterinarians' knowledge and attitudes in affected regions are not as advanced as needed, underscoring the importance of targeted epidemiological surveys on the knowledge of the disease.

Brucella canis and smooth Brucella species infections have been reported in dogs globally. In endemic countries such as Türkiye, dogs can be infected with both species. The exact incidence of canine brucellosis in Türkiye is unknown. This study aims to investigate the seroprevalence of brucellosis in stray dogs undergoing sterilization in Van province, eastern Türkiye. In this study, a comprehensive approach was used to ensure reliable incidence of Brucella spp in the dogs. We used the Rose Bengal Plate Test (RBPT) with both antigens from smooth Brucella abortus S99 (RBPT-S) and rough B. canis (RBPT-R) Brucella species to analyze 150 dog blood serum samples. For the in-house enzyme-linked immunosorbent assay (i-ELISA), we used a rough Lipopolysaccharide (LPS) antigen from the rough strain (i-ELISA-R) and an O-polysaccharide (O-PS) antigen (i-ELISA-S) from the smooth strain obtained from the OIE Reference Laboratory in Weybridge, UK. Each serum sample underwent analysis using a total of four serological tests, ensuring a thorough and reliable evaluation. Out of 150 serum samples analyzed, 32 (21.3 %) tested positive using the RBPT-S test and 12 (6.6 %) with the RBPT-R test. The i-ELISA-R identified 8 (5.3 %) positive samples, while the i-ELISA-S detected 27 (18 %) positive samples. The presence of B. canis and other Brucella spp. infections was detected in the dog in this region, revealing that canine brucellosis was primarily caused by smooth Brucella species. The test beased -smooth antigens identified a significantly higher number of positive samples compared to the test beased -rough antigens test, suggesting a potential difference in sensitivity or specificity between the two methods. This situation was considered an important risk for both dogs and humans, underscoring the urgent need for further research and the immediate implementation of public health measures to address this issue.

Canine leishmaniasis (CanL) caused by Leishmania infantum is a prevalent vector-borne disease in the Mediterranean region, notably Algeria. This research aims to assess the epidemiology and the clinical management practices of CanL employed by veterinarians in the Skikda and Constantine regions in the northeastern part of Algeria. A retrospective survey comprising 37 open and closed questions was distributed to 139 veterinary clinics between 2022 and 2023. The objective was to collect comparable epidemiological information on clinical presentations, diagnostic approaches, treatment protocols, preventive strategies, and awareness of the disease's public health implications. The findings from this study have revealed a notable prevalence of CanL in both regions, accompanied by similar clinical manifestations. Serological techniques, particularly rapid detection kits, are predominantly utilized alongside direct methods, while euthanasia remains the primary approach to combat the disease. The results show that only a few veterinarians educate pet owners about zoonotic diseases and prevention measures (32,79 %), while most do not provide this information. This investigation sheds more light on the current status of CanL in the selected studied area and suggests the need for standardizing diagnosis and treatment of CanL using evidence-based medicine criteria to enhance disease management. This will provide a proper way of conducting a more comprehensive epidemiological assessment of the condition for improving the handling and management of CanL in Algeria.

This study analyzed the molecular and phylogenetic profiles of Theileria annulata, the causative agent of tropical theileriosis in cattle, and its tick vector Hyalomma scupense in Algeria. Forty H. scupense ticks were collected in Medea, with 5 testing positive for Theileria spp. based on partial COXIII gene sequences. Positive ticks were further analyzed using COX1 and 12S rRNA genes. Two novel H. scupense 12S rRNA haplotypes and one novel COX1 haplotype were identified. One T. annulata haplotype previously reported in Algerian cattle was detected. This represents the first molecular characterization of T. annulata from H. scupense ticks in Algeria, providing insights into the genetic diversity of the parasite vector in this region. Overall, the study reveals new haplotypes for both the tick vector and parasite, furthering our understanding of their molecular profiles and phylogenetics in Algeria.

Cysticercosis, caused by larval stage of Taenia (T.) hydatigena is a disease of veterinary concern which causes major economic losses in livestock sector globally. Although livestock is the most productive part of Pakistan's economy, the epidemiological prevalence and genetic diversity of T. hydatigena has not been studied in great detail. Current study was undertaken for the estimation of the prevalence and genetic diversity of T. hydatigena among the goats and sheep of Multan in South Punjab, Pakistan. A PCR protocol targeting the cytochrome c oxidase subunit 1 (cox1) gene of T. hydatigena revealed an overall prevalence of 31.33 % among the small ruminants with higher prevalence in females (40.86 %) and older age group of >3-≤5 years (40.74 %). A total of 30 isolates were sequenced for partial mitochondrial cox1 gene which yielded 12 haplotypes with a haplotype diversity of 0.798 ± 0.68 and nucleotide diversity of 0.0046 ± 0.00183. A star like configuration was demonstrated in the haplotype network with a centrally positioned haplotype. Neutrality indices like Tajima's D and Fu's Fs were also biased towards negativity, a characteristic which suggests an expanding parasitic population. Moreover, the genetic diversity of T. hydatigena was also computed for the partial cox1 gene sequences available on GenBank, NCBI. A total of 415 sequences were retrieved out of which 142 haplotypes were identified having a haplotype diversity of 0.9150 ± 0.012 coupled with a nucleotide diversity of 0.00830 ± 0.00043. The haplotype profile of T. hydatigena population displayed an absence of a central haplotype, however, considerable genetic variation was ascertained. All the populations except Africa had a high haplotype diversity (>0.9), however, high Fst values were observed for a few populations which elucidated limited gene flow among the populations of Central/East Asia, Middle East, Europe and South Asia.

Hemoplasmas (HM) infect a wide range of mammals and may cause hemolytic anemia. We detected HM in a melon-headed whale (Peponocephala electra) stranded in Veracruz, Mexico. Sequencing and phylogenetic analysis of 16S and 23S rDNA indicated the presence of a HM that was previously detected in cetaceans from South America. This study expands the host and geographical range of detection of hemoplasmas in cetaceans from North America.

Dissemination of antimicrobial resistance in food animals is a One Health concern, but sheep production has been overlooked. This study aimed to explore the dissemination of Escherichia coli resistant to extended-spectrum cephalosporins (ESC) in feedlot lambs. Two pens were sampled on two occasions, and carcasses and other mammals living around were also tested. E. coli were recovered and antibiotic resistance determined. bla genes and their genetic localization were characterized. Whole genome sequencing (WGS) was performed to confirm clonal relationship. The most prevalent ESC-resistance genes in the 108 E. coli isolates were bla (53.7 %), bla (14.8 %) and bla (13.9 %). Most bla genes were found on the chromosome, but IncA/C, IncHI1, IncHI2 and IncF plasmids were also identified. Genetic diversity was observed even though ST6448 was by far the most frequent ST. WGS analysis showed high similarity among isolates recovered from feedlot lambs, animals in the surroundings and lambs' carcasses, proving the clonal and plasmidic dissemination.

Giardia intestinalis is a major diarrhea-causing parasite that colonizes the proximal small intestine of humans and various other mammalian species, including pets and livestock. Despite its global occurrence, there is limited information about the epidemiology of Giardia in reptiles, particularly snakes. The aim of this study was to amplify the beta-giardin (bg) gene of Giardia by polymerase chain reaction (PCR), followed by a comparative evolutionary tree analysis to determine the occurrence and molecular characterization of Giardia in snakes. We collected 603 asymptomatic samples from 26 provinces in China, representing species such as Pantherophis guttatus, Pantherophis obsoletus, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getula, and Heterodon nasicus. Ultimately, a occurrence of Giardia infection of 4.15 % was detected in these snakes, with corn snakes (Pantherophis guttatus) having a significantly higher occurrence than other species. Molecular analysis identified assemblage F as the predominant genotype, and also identified an assemblage B that can infect humans and a variety of mammals. The results of this study illustrate the potential risk of transmission of Giardia from snakes to humans, especially in environments where close contact occurs. The present epidemiological study examines epidemiological investigations of Giardia in reptiles, provides data to understand the zoonotic risk of Giardia, and emphasizes the need for targeted surveillance, stringent hygiene measures, and public awareness campaigns to reduce these risks.

Multi-drug resistant (MDR) Klebsiella pneumoniae is a public health concern due to its presence in Bangladeshi poultry products and its ability to spread resistance genes. This study genetically characterizes a distinct MDR K. pneumoniae isolate from the gut of poultry in Noakhali, Bangladesh, offering insights into its resistance mechanisms and public health impact.

Camel brucellosis is a zoonotic and economically important disease that causes low productivity and mortality in animals through abortion and low herd fertility.

Antimicrobial resistance (AMR) is fast emerging and is depleting antibiotics repertoire. Poultry is a major source for AMR because focus to enhance its production by modern practices widely uses antibiotics. India and China are major producers of meat and have hotspots of AMR. The Central and Southern India were predicted as emerging hotspots for AMR in poultry but no data available to substantiate it. To this end, we collected chicken feces from poultry farms in these regions and isolated genomic DNA. Further, shotgun whole genome sequencing was performed for metagenomics analysis. For the first time, we report the AMR gene profiles in poultry from Kerala and Telangana. The samples exhibited a higher prevalence of gram-negative and anaerobic species. The high priority pathogens in India were detected, like E.coli, Clostridium perfringens, Klebsiella pneumonia Staphylococcus aureus, Enterococcous faecalis, Pseudomonas aeruginosa, Bacteriodes fragiles. Conspicuously, the Southern India had the highest abundance of AMR genes than the Central India. E.coli was significantly more prevalent in the southernmost zone of India than in other sites. Our data had many common AMR profile features of the European Union (EU) poultry farms but lacked mcr-1, which was a lately emerged AMR gene in E.coli. Our data revealed the extent of AMR gene evolved in the Central and Southern India. It is comparable to the EU data but severity is lesser than in the EU.

Duck hepatitis A virus (DHAV) infection in ducklings causes acute hepatitis with considerable economic losses. In this study, Pekin and Muscovy duckling flocks (n=9) suffering from high mortality and hepatic lesions were examined by RT-PCR for DVHA. 44.4 % (4/9) of samples were positive for DHAV (5' UTR region), of which 100 % (4/4) were DVHA-3 (VP1 gene). VP1 sequencing and phylogenetic analysis of an isolate originated from Muscovy ducklings showed that it shared 96.8 % -100 %, 88.5-89.2 %, and 86.5-88.2 % nucleotide similarity (ns) with the Egyptian, Korean-Vietnamese, and Chinese DVHA-3 strains, respectively. It was distinguished from the DHAV-1 vaccine (67.6 % ns). The sequenced DVHA-3 isolate was used to experimentally infect 5-day-old Pekin and Muscovy ducklings vs. control groups. No apparent clinical signs or deaths were reported in the experimentally-infected groups. Pekin ducklings showed greater cloacal viral shedding than Muscovy until the 6th dpi (P<0.05). DVHA-3 induced a significant rise in IFN-β and IL-1β serum levels, where Muscovy ducklings' levels were higher than Pekin ducklings. Among the biochemical parameters, AST was only increased on the 4th dpi in both breeds vs. control (P<0.05). Compared to Muscovy ducklings at 2, 4, and 6 day post infection (dpi), the infected Pekin group had lower lipase levels (P≥0.05, p<0.05, and p<0.05, respectively), while ALT was higher at 4 and 6 dpi (P<0.05). The histopathological lesions supported the gross lesions, and their scores were dominant at 2 and 4 dpi in both breeds. At 6 and 8 dpi, Pekin showed more severe histopathological changes compared to Muscovy for the liver, heart, brain, and intestines; the pancreas, kidney, and lung showed the opposite pattern. In conclusion, Pekin ducklings displayed distinct DHAV-3 infection results from Muscovy ducklings, and more research utilizing a variety of DHAV-3 strains has to be carried out.

Bovine papular stomatitis (BPS) generally is a mild viral disease caused by the Bovine papular stomatitis virus (BPSV) belonging to the genus Parapoxvirus (PPV). This study aimed to perform the first molecular characterization and phylogenetic analysis of BPSV in beef calves in Iran. Clinical examinations were carried out on four beef cattle herds in West Azerbaijan province, which had experienced outbreaks of papular lesions. Fifty swab samples were collected from the papular lesions on the muzzle, lips, and oral cavity of affected calves, and after viral DNA extraction, they were subjected to polymerase chain reaction (PCR). The results of PCR confirmed the presence of BPSV in all calves with clinical symptoms. The partial B2L gene was sequenced based on nucleotides, and phylogenetic analysis was performed using MEGA 11.0.13. The analysis revealed that BPSV isolates from beef calves in Iran formed two clades. Clade 1 exhibited similarities to the isolates from Finland, Japan, and Georgia, while Clade 2 was similar to the Zambian isolates. These findings indicate the presence of genetic diversity and potential variability within the virus population. Further studies with larger sample sizes and diverse geographic regions will increase the resolution of the phylogenetic tree and contribute to a comprehensive understanding of how BPSV circulates in the country.