VETERINARY QUARTERLY

Circulating nucleosomes as a potential cancer biomarker in dogs with splenic nodular lesions
Meazzi S, Martini V, Marconato L, Aralla M, Licenziato L, Olimpo M, Roccabianca P, Sabattini S, Ubiali A, Zaccone R and Aresu L
Splenic nodular lesions in dogs can be either benign or malignant. They might be discovered incidentally or, in case of rupture, they may lead to hemoabdomen. Nevertheless, splenectomy followed by histopathology is essential for diagnosis and to prevent rupture. Yet, this invasive procedure might be postponed for dogs with benign splenic nodular lesions. Conversely, owners may opt for euthanasia over surgery for malignancies with poor prognosis like hemangiosarcoma. Thus, anticipating diagnosis with non-invasive biomarkers is crucial for proper patient management. In this prospective study, plasma samples were collected from 66 dogs with histologically confirmed splenic nodular lesions. A canine-specific ELISA kit was applied to assess nucleosome concentration, with histopathology of the spleen serving as the gold standard. Nucleosome concentration was found to be significantly higher in dogs with malignant splenic nodular lesions, particularly in those with hemangiosarcoma and other malignancies. The presence of hemoabdomen, more prevalent in dogs with splenic malignancy, also resulted in increased plasmatic nucleosome concentrations. Plasma nucleosomes could serve as a biomarker for detecting malignant splenic nodular lesions in dogs. More research is needed to understand how nucleosome concentration relate to disease stage and prognosis in dogs with hemangiosarcoma.
Transboundary viral diseases of pigs, poultry and ruminants in Southeast Asia: a systematic review
Mai TN, Nguyen TT, Dang-Xuan S, Nguyen-Viet H, Unger F and Lee HS
Livestock is a strategic part of the small-farm economy in Southeast Asia's society, providing food income, clothing, fertilizer, and draught power. However, incidences or outbreaks of transboundary animal diseases (TADs) are due to converging factors such as the natural hazards' aftermath, climate change, deforestation, urban growth, changing production conditions, and market chains. Therefore, this affects productivity and impacts farmers' livelihoods with small holdings. The literature review was carried out to understand the current situation of TADs in Southeast Asia, identifying knowledge gaps to provide actions for disease control and prevention in the region. We have attempted to summarise the scientific literature in English on the prevalence data of TADs in Southeast Asia between 2011 and March 2022. Relatively few studies evaluated the distribution of TAD, where most of the studies focused on diseases that are important for international trade, such as avian influenza (AI), African swine fever (ASF), classical swine fever (CSF), foot-and-mouth disease (FMD) and Newcastle disease (ND). Traditional production systems have received little attention in such studies as they belonged to mainly smallholders. The outbreaks of ASF and lumpy skin disease (LSD) in 2019 resulted in increased research activity between 2019-2022, while the other TADs were ignored in this period. For new emerging TADs diseases such as ASF and LSD, there is only information about the first detection without prevalence information. Therefore, further epidemiological investigations are necessary to reduce the gaps in disease surveillance reporting systems and support the prevention and reduction of further outbreaks.
Evaluation of the feasibility of equine in-vivo ultrasound technique for the medial branch of the dorsal ramus of the cervical spinal nerves
Nocera I, Di Franco C, Sorvillo B, Aliboni B, Bucchioni E, Sgorbini M, Sala G and Citi S
Ultrasound-guided local anaesthesia is commonly used in veterinary orthopaedics for horses. This study aimed to assess an ultrasound technique for the medial branch of the dorsal branch of the cervical spinal nerves (MB-DBCSNs) in horses and compare the performance of clinicians with different experience levels. Ten healthy, skeletally mature horses were examined using radiographic and ultrasound (US) techniques in the cervical area (C3-C7). Four operators with varying experience conducted US examinations using a 10 MHz linear and 6 MHz curvilinear transducer over ten training sessions. The number of cervical nerves visualized was recorded. A chi-square test was used to analyse the impact of training, anatomical location, and operator experience on the identification of facet joints. Operator agreement was evaluated with Cohen's K test. The operators assessed 80 MB-DBCSNs, with radiographs and identified 70 healthy and 10 pathological facet joints. Training significantly improved visualization success, reaching 90% in later sessions. Cranial facet joints (C3-C5) were more frequently visualized (81%) than caudal ones (C5-C7) were (59%). US performance was influenced by the operator's skill, and agreement among operators ranged from slight to fair. Overall, practice improved cervical nerve visualization , particularly for cranial nerves, but the technique requires a long learning curve because of low levels of operator agreement.
Pathological, immunological and molecular epidemiological analysis of lumpy skin disease virus in Indian cattle during a high-mortality epidemic
Manjunathareddy GB, Saminathan M, Sanjeevakumar L, Rao S, Dinesh M, Dhama K, Singh KP and Tripathi BN
Lumpy skin disease (LSD) is an economically significant, emerging viral disease of Cattle and Buffaloes. This study aimed to investigate the causes of high mortality in a recent LSD epidemic in India. We examined 1618 animals across seventy outbreaks and conducted post-mortem on 48 cattle out of 513 clinically suspected LSD cases. The morbidity, mortality and case fatality rates recorded were 31.70%, 2.97 and 9.37% respectively. Disease stages were categorized as early (20.81%), mid (42.02%), and late (37.17%) and the distribution of skin lesions was classified as mild (34.14%), moderate (39.39%), and severe (26.47%). Post-mortem findings revealed systemic infection with necrotic and ulcerative nodules on multiple internal organs. Histologically, necrotizing vasculitis and mononuclear cell infiltration with intracytoplasmic inclusions were observed in various organs. The highest viral load was found in skin nodules/scabs, trachea, tongue, and lymph nodes. The viral load was significantly higher in mid- and late-stages of skin nodules and internal organs; whereas, blood from early-stage showed high viral load. The expression of Th1-type and Th2-type cytokines varied significantly across different stages of the disease. The downregulation of the apoptotic intrinsic and upregulation of the extrinsic pathway genes, suggesting that the latter plays a role in LSDV infection. Genetic analysis revealed that the LSD virus (LSDV) isolates were derived from a Kenyan ancestral strain with unique nucleotide changes in RPO30 and P32 gene. In conclusion, the high mortality in the recent Indian LSD epidemic can be attributed to a newly identified, highly virulent strain of LSDV causing systemic infection.
A meta-analysis of livestock brucellosis prevalence in Indonesia
Ridlo MR, Andityas M, Primatika RA, Widantara H, Loong SK and Nuraini DM
Indonesia has a long history of livestock brucellosis, but the overall pooled prevalence remains unclear. This study aims to determine the pooled estimated prevalence of livestock brucellosis in Indonesia using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Five databases were searched and screened using predefined inclusion and exclusion criteria. Data from included studies were extracted and analyzed using a random effects model in R 4.3.0 for pooled estimated prevalence, subgroup meta-analysis, and meta-regression. Publication bias and sensitivity tests were conducted using Egger's test, funnel plot, trim and fill plot, and leave-one-out. The screening process identified 46 included studies, representing 47,057 samples for brucellosis testing. The pooled estimated prevalence for livestock brucellosis was 3.25% (95% CI, 1.81%-5.78%) with high heterogeneity ( = 2130.91,  = 0, =98%). Subgroup meta-analysis indicated no significant difference in the prevalence of livestock brucellosis across the main islands in Indonesia ( = 0.44) and across provinces in Sulawesi Island ( = 0.83), but significant differences were found among provinces in Java ( < 0.01). The subgroup meta-analysis based on animal type showed no significant difference between cattle, small ruminants, and pig brucellosis estimated prevalence ( = 0.26). Between serological tests, no significant difference was found ( = 0.77). Meta-regression showed no significant difference in brucellosis prevalence from 1988-2023. Egger's test and funnel plot showed publication bias. Trim and fill test indicated 21 studies should be added. As most studies were conducted in Java and Sulawesi Islands, caution should be exercised in interpreting the results, emphasizing the necessity of increasing the study of brucellosis in other regions.
The development of a lateral flow immunochromatographic test strip for measurement of specific IgA and IgG antibodies level against porcine epidemic diarrhea virus in pig milk
Jermsutjarit P, Venkateswaran D, Indrawattana N, Na Plord J, Tantituvanont A and Nilubol D
Porcine epidemic diarrhea virus (PEDV) causes severe enteric disease and high mortality in neonatal piglets, leading to significant economic losses in the swine industry. Considering that passive lactogenic immunity is crucial for preventing infection in piglets, necessitating a rapid and accurate tool to measure immunity levels. This study aims to develop a lateral flow immunochromatographic strip (LFICS) to assess IgA and IgG antibodies in colostrum and milk, using PEDV S protein. The performance of LFICS was compared to viral neutralization (VN) and enzyme-linked immunosorbent assay (ELISA) as reference methods, with a visual scoring system applied for field monitoring. Colostrum ( = 82) and milk ( = 106) samples were analyzed, showing strong correlation with reference methods and no cross-reactivity with other pig pathogens. The LFICS exhibited high relative sensitivity (Se) and specificity (Sp), with colostrum showing 98.73% Se and 66.67% Sp for IgA, and 96.15% Se and 75.00% Sp for IgG. Milk demonstrated 95.60% Se and 80.00% Sp for IgA, and 84.88% Se and 85.00% Sp for IgG. These findings indicate that the LFICS is a reliable, simple, and rapid method for measuring PEDV-specific IgA and IgG levels, offering valuable support for monitoring herd immunity and evaluating vaccination programs.
Polyomavirus surveillance in cetaceans of Brazil: first detection of polyomavirus in Guiana dolphins ()
Duarte-Benvenuto A, Díaz-Delgado J, Ewbank AC, Groch KR, Dal Bianco V, Costa-Silva S, Zamana-Ramblas R, Favero C, Castaldo Colosio A, da Cunha Gomes Ramos H, Santos-Neto E, Lailson-Brito J, Carvalho VL, Ribeiro VL, Pacheco Bertozzi C, F Pessi C, Sacristán I, Catão-Dias JL and Sacristán C
Polyomaviruses (PyVs) are small double-stranded DNA viruses able to infect species across all vertebrate taxa. In cetaceans, PyVs have been reported only in short-beaked common dolphin (), common bottlenose dolphin () and killer whale (). Herein, we surveyed PyV in 119 cetaceans (29 mysticetes and 90 odontocetes) stranded along the Brazilian coast, from 2002 to 2022, comprising 18 species. DNA extracted from the lungs was tested using a nested PCR targeting the major capsid protein gene of PyV. Polyomavirus was detected in lung samples of 1.7% (2/119) cetaceans: two juvenile female Guiana dolphins () stranded in Rio de Janeiro (Rio de Janeiro state) and Guriri (Espírito Santo state), in 2018. Both retrieved sequences were identical and presented 93.3% amino acid identity with , suggesting a novel species. On histopathology, one of the PyV-positive individuals presented basophilic intranuclear inclusion bodies morphologically consistent with polyomavirus in the lungs. Other available tissues from both cases were PyV-PCR-negative; however, both individuals tested positive for Guiana dolphin morbillivirus. To our knowledge, this is the first report of PyV infection in cetaceans of the Southern Hemisphere and the first description of a co-infection with morbillivirus.
Pharmacokinetics, optimal dosages and withdrawal time of amoxicillin in Nile tilapia () reared at 25 and 30 °C
Rairat T, Lu YP, Ho WC, Ke HJ and Chou CC
Knowledge of amoxicillin (AMX) pharmacokinetics (PK) and tissue residues in fish, which is necessary for prudent drug use, remains limited. The study aimed to explore the PK characteristics of AMX in Nile tilapia () reared at 25 and 30 °C as well as to determine optimal dosages and drug withdrawal time (WDT). In the PK investigation, the fish received a single dose of 40 mg/kg AMX oral gavage, and the optimal dosage was determined by the pharmacokinetic-pharmacodynamic approach. In the tissue residue study, the fish were orally gavaged with 40 mg/kg/day AMX once daily for 5 days and the WDT was established by the linear regression analysis. The results revealed the temperature-dependent drug elimination; the clearance relative to bioavailability (CL/F) and elimination half-life at 30 °C (0.180 L/kg/h and 6.06 h, respectively) were about twice those at 25 °C (0.090 L/kg/h and 10.49 h, respectively). The optimal dosages at the minimum inhibitory concentration (MIC) of 2 μg/mL were 10.97 (25 °C) and 41.03 (30 °C) mg/kg/day, respectively. Finally, following the multiple oral administration, the muscle/skin residue of AMX on day 1 after the last dosing at 25 and 30 °C were 548 and 264 ng/g, respectively. The average tissue residues were depleted below the maximum residue limits (MRL) of 50 μg/kg on day 5 (25 °C) and 3 (30 °C), respectively, and the WDT were 6 and 4 days when rearing at 25 and 30 °C, respectively. This knowledge serves as a practical guideline for responsible use of AMX in treating bacterial diseases in Nile tilapia aquaculture.
Advances research in porcine enteric coronavirus therapies and antiviral drugs
Liang J, Xu W, Pan X, Han S, Zhang L, Wen H, Ding M, Zhang W and Peng D
The porcine enteric coronaviruses (PECs) currently reported include porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV), transmissible gastroenteritis virus (TGEV), and swine acute diarrhea syndrome coronavirus (SADS-CoV). In the absence of effective treatment, they can cause similar clinical characteristics including weight loss, sleepiness, vomiting, anorexia and fatal diarrhea in neonatal piglets, resulting in significant economic losses to the global pig industry. Although many studies on drugs for treating and combating PECs have been issued. There are still no specific drug targeting PECs and used in clinical production. Therefore, it is necessary to sort out and summarize the research on the treatment and anti PECs drugs, and further development of low toxicity and high efficiency drugs is needed. Here, we review the latest progress of anti PECs drugs, focus on the mechanism of anti PECs reaction of drug components, and try to clarify new strategies for effective control and elimination of PECs. These comprehensive and profound insights will help to further investigate, prevent and control the transmission of PECs infection.
Beclin-1 and LC3B expression in canine mast cell tumours: an immuno-ultrastructural and immunohistochemical study of autophagy
Vicente GP, Della Salda L and Strefezzi RF
Mast cell tumours (MCTs) are common malignant neoplasms in dogs, for which prognosis and therapeutic decisions are based on histological features and proliferation markers. Autophagy is a cellular catabolic process responsible for degrading cytoplasmic components to maintain homeostasis, alterations in which are frequently linked to tumour growth and progression. This study was conducted to investigate the occurrence of autophagy in canine MCTs and to verify its value as a prognostic indicator for dogs with the disease. Beclin-1 and LC3B expressions were investigated using immunohistochemistry, and autophagy was ultrastructurally characterised. The autophagic phenomenon was successfully visualised in neoplastic mast cells under transmission electron and immunoelectron microscopy. MCTs from dogs that died due to the disease showed higher positivity for Beclin-1 and dogs with MCTs presenting a LC3B granular immunohistochemical pattern had a significantly shorter post-surgical survival. The occurrence of autophagy is an indicator of poor prognosis. Future studies are needed to elucidate the specific mechanisms and open new opportunities to treatments targeting this cancer cell advantage.
Assessment of the exposure to and the presence of anti- antibodies in stray cats in an endemic region of Spain, and their potential correlation with environmental factors
Marteles D, Martínez MV, Fernández A, Riera C, Fisa R, Roca-Geronès X, Chavez-Fisa S, Castañeda S, Ramírez JD, Davis JE, Sumova P, Volf P, Verde M, González A, Alcover MM and Villanueva-Saz S
is a major vector of in the Mediterranean. While the seroprevalence of leishmaniosis in Spanish dogs and cats has been studied, data on the exposure of cats to bites under natural conditions without repellents is limited. Stray cats could serve as sentinels for and exposure. This study analyzed sera from 204 apparently healthy stray cats, collected from January 2021 to January 2022, for antibodies against saliva and parasites. Anti-sand fly antibodies were detected in 40.69% of cats using an ELISA with the recombinant salivary protein SP03B of . Seroprevalence of infection was 23.52% by Western blot and 27.41% by ELISA, with an overall seroprevalence of 40.69% (95% CI 34.18-47.54%). This is the first assessment of antibody response to saliva and in naturally exposed stray cats in Spain. Further research is needed to examine the salivary antigens recognized by cats and to explore the relationship between exposure and infection severity in cats.
New preclinical biomarkers for prion diseases in the cerebrospinal fluid proteome revealed by mass spectrometry
Pérez-Lázaro S, Barrio T, Bravo SB, Sevilla E, Otero A, Chantada-Vázquez MDP, Martín-Burriel I, Requena JR, Badiola JJ and Bolea R
Current diagnostic methods for prion diseases only work in late stages of the disease when neurodegeneration is irreversible. Therefore, biomarkers that can detect the disease before the onset of clinical symptoms are necessary. High-throughput discovery proteomics is of great interest in the search for such molecules. Here we used mass spectrometry to analyse the cerebrospinal fluid proteome in an animal prion disease: preclinical and clinical sheep affected with natural scrapie, and healthy sheep. Interestingly, we found 46 proteins in the preclinical stage that were significantly altered ( < 0.01) compared to healthy sheep, mainly associated with biological processes such as stress and inflammatory responses. Five of them were selected for validation by enzyme-like immunosorbent assay: synaptotagmin binding, cytoplasmic RNA interacting protein (SYNCRIP), involved in nucleic acid metabolism; phospholipase D3 (PLD3) and cathepsin D (CTSD), both related to lysosomal apoptosis; complement component 4 (C4), an element of the classical immune response; and osteopontin (SPP1), a proinflammatory cytokine. These proteins significantly increased in the preclinical stage and maintained their levels in the clinical phase, except for CTSD, whose concentration returned to basal levels in the clinical group. Further research is ongoing to explore their potential as preclinical biomarkers of prion diseases.
A high titer antibody response against P22 protein immunocomplex is not correlated with protection in naturally tuberculosis-infected goats
J O, I AR, A R, I M, A GB, B R, E FC, L J, M D, L D, M A R and J B
Caprine livestock are significant reservoirs of the complex (MTBC), contributing to tuberculosis (TB) transmission among animals and humans. The P22 protein immunocomplex (P22PI), derived from bovine tuberculin, shows immunostimulating capacity and is used for TB diagnosis. This study assessed the immunogenicity and protective efficacy of P22PI in two groups of goats: 24 naïve goats (12 immunised, 12 controls) from a TB-free herd, and 24 infected goats (12 immunised, 12 controls), referred to as pre-infected animals, from a -infected herd. Both were exposed for 5 months to -naturally infected goats. Reactors to single and comparative intradermal tuberculin (SIT and SCIT, respectively) tests and interferon-gamma release assay (IGRA) significantly increased ( < 0.05) in both groups 5 months' post-exposure, with no significant differences between immunised and control animals. However, immunised animals exhibited a significantly higher ( < 0.05) antibody response against P22PI. Most naïve animals (83.3%) and all pre-infected animals developed TB-compatible lesions, with extensive necrosis in the lungs and associated lymph nodes, compared to 50% and 83.3% of control animals, respectively. These findings suggest that while P22PI stimulates an intense antibody response under the conditions of the present study, it does not confer protection against TB and may exacerbate disease severity.
Seroprevalence and risk factors of bluetongue virus in domestic cattle, sheep, goats and camels in Africa: a systematic review and meta-analysis
Medrouh B, Abdelli A, Belkessa S, Ouinten Y, Brahimi M, Hakem A, Kernif T, Singer SM, Ziam H, Tsaousis AD, Jokelainen P, Savini G and Pasolli E
Bluetongue (BT) is a vector-borne disease affecting wild and domestic ruminants in many parts of the world. Although bluetongue virus (BTV) is widespread in ungulates in Africa, available epidemiological information on BT in this continent is limited. This systematic review and meta-analysis aimed to estimate the seroprevalence of BTV and summarize information on associated risk factors in domestic ruminants and camels in Africa. Systematic searches were conducted from the inception of the database to November 2022 on PubMed/MEDLINE, ScienceDirect, Web of Science, and Google/Google Scholar. Forty-four eligible publications were identified, published in the range from 1973 to 2020, and statistically analyzed. The pooled overall seroprevalence of BTV was 45.02% (95% confidence interval [CI]: 36.00-54.00%). The pooled seroprevalence was 49.70% (95% CI: 34.50-65.00%) in cattle, 47.00% (95% CI: 29.90-64.50%) in goats, 40.80% (95% CI: 19.60-63.90%) in camels, and 36.30% (95% CI: 29.00-44.90%) in sheep. The pooled seroprevalence decreased after 1990 and increased again after 2010. The highest pooled overall seroprevalence was found in the southeastern region, and the highest pooled overall seroprevalence was obtained by Competitive Enzyme-Linked Immunosorbent Assay. Finally, the seroprevalence in females (53.30%, 95% CI: 34.80-71.00%) was significantly higher than in males (28.10%, 95% CI: 17.40-40.30%) ( < 0.05). We showed that antibodies against BTV were common in African ruminants and camels. Monitoring the seroprevalence of BTV, as well as systematic and continuous surveillance of the population, are encouraged to prevent and control the spread of BT.
Molecular identification and biological characterization of from domestic pigeons () in Guangdong, China
He X, Fang XM, Qiao YT, Su JL, Tang SQ, Weng YB and Lin RQ
Pigeon coccidiosis caused by spp. is an important veterinary disease with a significant economic impact on the pigeon industry. Preventive measures for in pigeons have been hampered by the lack of extensive genetic, morphological, and biological data on the oocysts. In this study, we examined the prevalence and identity of spp. in domestic pigeons from seven cities in Guangdong Province, China. Data show that coccidiosis was prevalent in domestic pigeons in Guangdong Province, with an overall spp. detection rate of 73.4%. Five species were identified, including (73.4%), (25.6%), (19.6%), (19.6%), and (6.7%). We obtained single oocyst-derived lines of the dominant from fecal specimens. oocysts measured 20.06 ± 0.69 μm × 18.63 ± 1.03 μm, and sporocysts measured 10.29 ± 0.82 μm × 85.38 ± 0.46 μm. In infection experiment using obtained isolates, 60-day-old coccidia-free pigeons exhibited a prepatent period of 105 h and patent period of 9-10 days followed by severe diarrhea, depression, anorexia, and emaciation. Endogenous development of the parasite was observed mainly in the cytoplasm of epithelial cells in the duodenum, jejunum, ileum, and rectum. Two generations of meronts developed on days 3 and 4 after infection, respectively, while gamont and gamete developed on day 5 after infection. The morphological, genetic, and biological data are expected to be useful in elucidating the biological characterization of pigeon coccidiosis to develop measures against the treatment and containment of this disease.
The global seroprevalence of infection in workers occupationally exposed to animals (1972-2023): a systematic review and meta-analysis
Mohammed A, Ahmed M and Ibrahim N
, a ubiquitous zoonotic parasite infecting warm-blooded animals, poses a significant health threat to workers with occupational animal exposure (WOEA) due to their frequent contact with potential reservoirs. Existing data on seroprevalence in the WOEA exhibits substantial global variation. This systematic review and meta-analysis, adhering to PRISMA guidelines, aimed to quantify the global seroprevalence of infection among WOEA over the past five decades (1972-2023). We identified 66 eligible studies through a comprehensive search strategy encompassing English publications, with a total sample size of 15,279. A random-effects model with the Freeman-Tukey transformation in STATA v16.0 accounted for the high heterogeneity observed. We estimated the pooled global seroprevalence of infection in WOEA at 41% (95% CI: 36-47%). Subgroup analyses revealed significant variations by gender: males (63%) vs. females (37%) ( < 0.05), occupation: non-livestock workers (54%), livestock workers (47%), slaughterhouse workers (44%), and veterinary personnel (27%) ( < 0.05). Geographic trends showed the highest prevalence in Africa (51%), followed by South America (49%), Europe (47%), Australia (43%), Asia (36%), and North America (23%;  < 0.05). Lower prevalence was observed in high-income (39%) and upper-middle-income (38%) countries compared to lower-middle-income (44%) and low-income (48%) countries ( < 0.05). This analysis underscores the high global seroprevalence of in the WOEA, highlighting the need for targeted interventions in this high-risk population.
Canine parvovirus in North-East India: a phylogenetic and evolutionary analysis
Jayappa K, Rajkhowa TK and Gaikwad SS
Canine parvovirus type 2 (CPV-2) infection in dogs is considered as one of the most common cause of morbidity and mortality in young dogs and continues to occur with high incidence worldwide. Despite a single-stranded DNA virus, CPV-2 possesses a high mutation rate which has led to the development of new variants from time to time. These variants are classically classified based on the amino acid markers present in the VP2 gene. In this study, we examined 20 different cases of CPV-2 infection from seven different states of the North East region (NER) of India. The near-complete genome sequences of all these isolates were subjected to phylodynamic and phylogeographic analysis to evaluate the genetic diversity and geographical spread of CPV-2 variants. Analysis of the deduced amino acid sequences revealed residues characteristic of the 'Asian CPV-2c lineage' in all the 20 sequences confirming it as the dominant strain circulating in NER, India. The phylogenetic analysis based on the whole genome showed that all 20 sequences formed a monophyletic clade together with other Asian CPV-2c sequences. Furthermore, phylogeographic analysis based on the VP2 gene showed the likely introduction of Asian CPV-2c strain to India from China. This study marks the first comprehensive report elucidating the molecular epidemiology of CPV-2 in India.
TGFβ in malignant canine mammary tumors: relation with angiogenesis, immunologic markers and prognostic role
Carvalho MI, Silva-Carvalho R, Prada J, Pinto C, Gregório H, Lobo L, Pires I and Queiroga FL
Transforming growth factor-β (TGFβ) and FoxP3 regulatory T cells (Treg) are involved in human breast carcinogenesis. This topic is not well documented in canine mammary tumors (CMT). In this work, the tumoral TGFβ expression was assessed by immunohistochemistry in 67 malignant CMT and its correlation to previously determined FoxP3, VEGF, and CD31 markers and other clinicopathologic parameters was evaluated. The high levels of TGFβ were statistically significantly associated with skin ulceration, tumor necrosis, high histological grade of malignancy (HGM), presence of neoplastic intravascular emboli and presence of lymph node metastases. The observed levels of TGFβ were positively correlated with intratumoral FoxP3 (strong correlation), VEGF (weak correlation) and CD31 (moderate correlation). Tumors that presented a concurrent high expression of TGFβ/FoxP3, TGFβ/VEGF, and TGFβ/CD31 markers were statistically significantly associated with parameters of tumor malignancy (high HGM, presence of vascular emboli and nodal metastasis). Additionally, shorter overall survival (OS) time was statistically significantly associated with tumors with an abundant TGFβ expression and with concurrent high expression of TGFβ/FoxP3, TGFβ/VEGF, and TGFβ/CD31. The presence of lymph node metastasis increased 11 times the risk of disease-related death, arising as an independent predictor of poor prognosis in the multivariable analysis. In conclusion, TGFβ and Treg cells seem involved in tumor progression emerging as potential therapeutic targets for future immunotherapy studies.
Predictive value of RNAscope hybridization and p53 immunohistochemistry for mutational status in canine diffuse large B-cell lymphoma
Foiani G, Licenziato L, Marconato L, Fanelli A, Melchiotti E, Zanardello C, Aresu L and Vascellari M
mutations are associated with short survival and poor treatment response in canine diffuse large B-cell lymphoma (cDLBCL). The expression of by RNAscope hybridization and p53 by immunohistochemistry (IHC) was investigated in 37 formalin-fixed paraffin-embedded cDLBCL, to assess their correlation with mutational status and to evaluate their prognostic value. was detected in all samples by RNAscope. Ten of 37 (27%) cases expressed p53 by IHC, with highly variable percentage of positive cells. RNAscope scores and p53 IHC results were not correlated. The expression of by RNAscope was not influenced by its mutational status. Conversely, p53 IHC and mutations were significantly associated. p53 IHC predicted genetic mutations with high accuracy (97.3%). All mutated samples carrying missense mutations exhibited p53 expression by IHC, while all wild-type cases and a single case with frameshift insertion were negative. In univariable analysis, p53 IHC was associated with shorter time to progression (TTP) and lymphoma-specific survival (LSS). Nevertheless, in multivariable analysis, only treatment significantly affected TTP and LSS. These findings suggest p53 IHC is an accurate, cost-effective tool for predicting mutations in cDLBCL, unlike RNAscope, though its prognostic value requires further validation.
Detection and quantification of DNA in water buffaloes () using droplet digital polymerase chain reaction
Fusco G, Cardillo L, Valvini O, Pucciarelli A, Picazio G, Cerrone A, Napoletano M, Pellicanò R, Ottaiano M, de Martinis C, De Falco F, Cutarelli A, Sannino E, Borriello G, Tittarelli M, Roperto S and De Carlo E
Brucellosis represents a major public health concern worldwide. Human transmission is mainly due to the consumption of unpasteurized milk and dairy products of infected animals. The gold standard for the diagnosis of spp in ruminants is the bacterial isolation, but it is time-consuming. Polymerase Chain Reaction (PCR) is a quicker and more sensitive technique than bacterial culture. Droplet digital PCR (ddPCR) is a novel molecular assay showing high sensitivity in samples with low amount of DNA and lower susceptibility to amplification inhibitors. Present study aimed to develop a ddPCR protocol for the detection of in buffalo tissue samples. The protocol was validated using proficiency test samples for spp by real time qPCR. Furthermore, 599 tissue samples were examined. Among reference materials, qPCR and ddPCR demonstrated same performance and were able to detect up to 225 CFU/mL. Among field samples, ddPCR showed higher sensitivity (100%), specificity and accuracy of 93.4% and 94.15%, respectively. ddPCR could be considered a promising technique to detect in veterinary specimens, frequently characterized by low amount of bacteria, high diversity in matrices and species and poor storage conditions.
Comparison of different dietary levels of iron and zinc for the enrichment of eggs in laying hens
Ullah Z, Khan S, Shuaib M, Ullah O, Siddiqui SA, Pokoo-Aikins A, Jabir MS and Swelum AA
Zinc (Zn) and iron (Fe) are necessary for structural and metabolic functions of the body like, immune competence, physical growth, reproductive function, and neurobehavioral development. The deficiency of micronutrients is a severe public health distress in nearly all developing countries which leads to the syndrome of malnutrition mostly in children and women. The present study was conducted to compare the effect of different levels of Fe and Zn on feed intake (FI), feed conversion ratio (FCR), daily egg production, and Fe and Zn depositions in the egg yolk of laying hens. For this purpose, 80 commercial laying birds were allocated into four groups consisting of four replicates each ( = 5). The basal ration was provided to the control group (C) while three different levels (100, 200, and 300 mg/kg diet/element) of Fe and Zn (1:1) were provided to the FZ100, FZ200, and FZ300 groups, respectively. Overall FI, hen day egg production, and FCR were not affected ( > 0.05). The Fe and Zn content in egg yolk were significantly increased in the FZ300 group after 4 weeks of supplementation. The cost per egg produced in response to different levels of Fe and Zn supplementation was not affected however a numerical increase in cost/egg was found in the FZ300 group. In conclusion, Fe and Zn supplementation (300 mg/kg diet/element) significantly enhanced the egg Fe and Zn contents without adversely affecting production parameters including FI, egg production, and FCR of laying hens.