This study evaluated the in vitro probiotic-related properties and preliminary safety profiles of five lactic acid bacteria (LAB) strains for potential use in aquaculture: Limosilactobacillus reuteri ATCC 55730, Lentilactobacillus kefiri K107, Latilactobacillus sakei SN34, Lactiplantibacillus plantarum FI8595, and Streptococcus thermophilus BioSt64. Probiotic-related traits included antimicrobial activity against Lactococcus garvieae and Yersinia ruckeri, aggregation-related properties, and tolerance to fish bile and acidic conditions. Safety assessment included hemolytic and enzymatic activities and antibiotic susceptibility. Lmb. reuteri showed the strongest inhibition of L. garvieae (18.25 ± 1.06 mm), followed by S. thermophilus (9.85 ± 0.21 mm), whereas Ltb. sakei inhibited Y. ruckeri (9.00 ± 0.00 mm). All strains coaggregated with both pathogens and remained viable under the tested acidic and bile conditions. Lmb. reuteri and Lpb. plantarum showed high autoaggregation and cell surface hydrophobicity, which may indicate potential for intestinal adhesion, although this should be confirmed through direct adhesion assays. S. thermophilus showed the most favorable preliminary profile, combining antimicrobial activity, absence of detectable hemolysis, and susceptibility to most antibiotics under the conditions tested. Lmb. reuteri also showed promising probiotic-related properties, but its previously reported antimicrobial resistance determinants represent an important safety concern, while the β-hemolytic activity of Ltb. sakei precludes its selection without further assessment. Further controlled in vivo evaluation is required before any of these strains can be considered for use as aquaculture probiotics.

Escherichia coli (E. coli) is a common cause of bovine mastitis, and rapid detection from milk is important for timely disease management. However, molecular assays such as colourimetric loop-mediated isothermal amplification (LAMP) are often limited by laboratory-based DNA extraction. This study evaluated a simplified HotSHOT (HS) DNA preparation method for extracting E. coli DNA from bovine milk and assessed its compatibility with colourimetric LAMP and conventional PCR. Pasteurised bovine milk spiked with E. coli was used for analytical evaluation, and culture‑positive clinical milk samples were tested to assess diagnostic performance. The HS method was compared with five commercial DNA extraction kits and a direct boiling method without centrifugation. Performance was assessed using limit of detection (LOD) expressed as colony-forming units per millilitre. Using the HS method, the LOD was 7.9 × 10[4] CFU/mL for LAMP and 7.9 × 10[3] CFU/mL for PCR. PCR performance of the HS method was comparable to the QIAamp and Norgen commercial kits, while LAMP sensitivity was ten- to a hundred-fold lower than the best kit-based methods. Direct boiling showed markedly reduced sensitivity, with an LOD of 7.9 × 10[8] CFU/mL. Among 17 culture-confirmed clinical mastitis milk samples positive for E. coli, LAMP detected 15 (88.2%) and PCR detected all 17 (100%). These results show that simplified DNA preparation can support molecular detection of E. coli in bovine milk and provide a practical basis for future development of field-deployable diagnostics in resource-constrained settings.

One of the most important bacterial agents of colibacillosis is Avian Pathogenic Escherichia coli (APEC), which affects the poultry industry by increasing mortality, reducing growth performance, and ultimately decreasing production efficiency. According the surveillance studies carried out in the European Union during the period of 2019-21, resistance to antibiotics has increased in E. coli due to transfer of genes between bacteria and humans. The rapid emergence of high antimicrobial resistance has accelerated research towards effective, natural, and antibiotic-free substitutes. Curcumin is a bioactive polyphenol extracted from Curcuma longa, has been widely studied for its antioxidant, immunomodulatory, antimicrobial, and anti-inflammatory properties. But, its use as a feed additive for chickens is limited because of the low bioavailability, rapid metabolism, and poor solubility. By applying nanotechnology techniques, researchers have successfully developed formulations that counteract these disadvantages and improve their bioavailability and stability. Curcumin nanoparticles work by causing damage to bacterial cell membrane, production of reactive oxygen species, and impeding bacterial cytokinesis by preventing the assembly of the FtsZ protein. Applications of nanoformulations of curcumin as a therapeutic agent for increasing the production of chicken, the various methods of delivering them to the birds, their pharmacokinetic studies and their effect on the growth performance of poultry birds have been critically and thoroughly analyzed in this review. Nanocurcumin at a concentration of 100 mg per kilogram of feed has demonstrated the best results for body weight gain, feed conversion ratio and feed intake. Despite the positive results, there are unsolved queries about the toxicokinetics, signaling molecular pathways, long-term safety, and dose standardization. For nanocurcumin to be logically incorporated into chicken health management, these obstacles must be overcome.

This study investigated the antimicrobial activity of rosemary essential oil (REO) as a natural alternative to conventional disinfectants for Japanese quail hatching egg sanitization, focusing on the differential susceptibility of presumptive cultivable bacterial groups associated with eggshell contamination. Following previous findings demonstrating that REO reduces total microbial load and improves hatchability and chick performance, the present study aimed to characterize the response of specific cultivable bacterial groups to REO treatment. Eggs were divided into four groups: untreated control (C-), 70% alcohol-treated (C[+ A]), quaternary ammonium-treated (C[+ Q]), and REO-treated at 10 µL/mL. Cultivable bacterial populations were assessed using serial dilutions and selective culture media targeting specific Gram-positive and Gram-negative bacterial groups. Presumptive identification based on culture and morphological characteristics suggested the presence of Lactobacillus spp., Bacillus spp., Acinetobacter spp., Staphylococcus spp., Escherichia coli, and Enterobacter spp. REO treatment reduced several cultivable eggshell bacterial groups, with the greatest reduction observed for presumptive Escherichia coli, whereas presumptive Lactobacillus spp. were comparatively less affected. The response to REO therefore varied among the bacterial groups evaluated. In comparison, conventional sanitizers produced greater reductions in some bacterial groups, particularly presumptive Lactobacillus spp. These findings indicate that REO exhibits a differential antimicrobial profile against cultivable eggshell bacterial populations and highlight its potential as a natural alternative for hatching egg sanitization. Under the conditions investigated, the 10 µL/mL concentration may provide a favorable balance between antimicrobial activity and embryonic safety.

Epsilon toxin (Etx) is a major exotoxin secreted by Clostridium perfringens that causes Enterotoxaemia (ET) in small ruminants leading to 'sudden death' and is known to induce protective antibodies. This study describes an indirect-ELISA based on recombinant quadruple-point Etx-mutant protein (~ 36 kDa), a well-defined and non-toxic antigen to detect epsilon specific antibodies in sheep and goat sera samples. The assay was evaluated for diagnostic sensitivity and specificity, along with cross-reactivity, as well as compared with a commercial ELISA kit. Furthermore, rEtx-iELISA was used to assess antibody titers in randomly collected sheep (n = 584) and goat (n = 555) serum samples from different regions of Odisha, an ET endemic state in India. The rEtx mutant protein (125 ng/well) based indirect ELISA platform demonstrated the area under the curve (AUC) of 0.997, diagnostic sensitivity (DSn) of 98.7%, and diagnostic specificity (DSp) of 96.2% using a pre-established test serum panel with 95% CI of 93.2-99.2% (p < 0.0001) against an optimal cut-off criterion of 10.42 PP. A comparison of the assay with a commercial kit yielded a kappa value of 0.912. A higher (> 95%) degree of sero-positivity with an average protective flock immunity of > 56% (≥0.15 IU/mL) was noted in sheep and goat populations of Odisha state. The study indicated the potential diagnostic utility of recombinant Etx-mutant antigen based indirect-ELISA for sero-monitoring of ET-specific antibodies in small ruminants.

Amyloidosis is a disease characterized by the deposition of abnormally folded proteins as amyloid fibrils in tissues and occurs in various mammals and birds. In some cases, amyloids derived from two different precursor proteins may coexist in the same individual, a condition known as double amyloidosis. Amyloid A (AA) amyloidosis is known to occur frequently in birds; however, little is known about other types of amyloidosis. In the present study, we report a case of amyloidosis in a Humboldt penguin involving two distinct types of amyloids. Histopathological examination, transmission electron microscopy, mass spectrometry-based proteomic analysis, and immunohistochemistry revealed the concurrent occurrence of fibrinogen Aα-chain (FibA) amyloidosis and AA amyloidosis. FibA-derived and AA amyloid deposits exhibited distinct nodular and diffuse morphologies, respectively, suggesting that their deposition patterns may provide a morphological clue for distinguishing between the two types of amyloids. To our knowledge, this is the first case of FibA amyloidosis in a penguin and the first report of the concurrent occurrence of two distinct types of systemic amyloidosis in a bird.

Phages show promising application prospects as antibacterial agents for the treatment of drug-resistant bacterial infections. However, the rapid evolution of phage resistance in bacteria has compromised the therapeutic efficacy. Here, we first screened a broad-host-range phage PC13 based on the Salmonella-phage library. Then, phage-resistant Salmonella mutants were obtained by serial co-culture passages of PC13 Salmonella enterica subsp. enterica serovar Enteritidis C2. Whole-genome sequencing analyses were performed on the phages and resistant strains to decipher the potential molecular mechanisms of phage-host interactions and collateral sensitivity. The bacteriostatic and therapeutic effects of the triple phage cocktail (PC13 + PC11+PC16) against S. Enteritidis C2 were evaluated using in vitro bacteriostatic assays and in vivo chicken infection models. The results showed that PC13 lysed 90% (81/90) of the tested chicken-derived Salmonella isolates. PC11 achieved a lysis rate of 95% against all 20 PC13-resistant mutants. Relative to single-phage PC13 and the PC13 + PC11 cocktail, the triple-phage cocktail PC13 + PC11 + PC16 markedly extended bacteriostatic time in vitro (p < 0.01). The phage cocktail significantly prolonged the bacteriostatic time in vitro compared to individual phages (p < 0.01). Comparative genomic analysis revealed that 95% of the PC13-resistant mutants harbored missense, nonsense, or frameshift mutations in genes associated with lipopolysaccharide (LPS) core oligosaccharide synthesis. These PC13-resistant mutants exhibited significantly enhanced susceptibility to PC11 and PC16 relative to the wild-type strain (p < 0.05). We propose that Salmonella evaded PC13 infection by altering LPS structure via these mutations, and these genetic changes simultaneously exposed the putative binding receptors predicted for PC11 and PC16, conferring collateral sensitivity of the resistant mutants to the latter two phages. In the chicken S. Enteritidis infection model, compared with the Salmonella treatment group, the fecal Salmonella load of chicks in the cocktail treatment group decreased by about 4.0 log CFU/g, and there was no significant difference in body weight changes compared with control group. In conclusion, a phage cocktail against S. Enteritidis with significantly enhanced antibacterial activity was rationally formulated constructed by targeting phage-resistant escape mutants generated during phage-bacterial coevolution. The collateral sensitivity effect uncovered in this study explains its superior inhibitory performance, which provides theoretical and practical strategies for developing highly efficient phage therapy.

Listeria monocytogenes is a zoonotic bacterium widely distributed in the environment and capable of infecting a broad range of animal species, including humans, usually as a food-borne disease. Although listeriosis is well recognized across multiple hosts, information in small mammals such as rodents and lagomorphs remains comparatively limited, with most reports describing sporadic systemic disease. This study describes two cases of septicemic listeriosis with reproductive involvement in pregnant long-tailed chinchillas (Chinchilla laniger) from a breeding facility in Argentina. Both animals developed depression, anorexia, prolonged gestation, and died. Grossly, both animals had mucosanguineous vaginal discharge and fetal maceration; one also showed multifocal random hepatic necrosis. Histopathology revealed severe multifocal necrosuppurative hepatitis, enteritis, endometritis and placentitis, and pulmonary thromboembolism. Immunohistochemistry demonstrated abundant intralesional Listeria spp. antigen in the liver and uterus. L. monocytogenes DNA was detected in the liver by real-time PCR. The present work expands the knowledge of listeriosis in chinchillas by providing detailed documentation of systemic infection with reproductive involvement. Given the zoonotic nature of L. monocytogenes, early recognition of infection in captive animal populations is relevant for both animal and public health.

This case report describes the management of a complex volumetric muscle loss (VML) injury in a 3-year-old mixed-breed dog following severe trauma to the right forelimb, which resulted in extensive multi-muscle defects that were unsuitable for primary closure. After initial stabilization and surgical debridement, reconstruction was achieved using a transpositional flap, while a gold nanoparticles-bioactive glass-alginate-cellulose (BGAuSP-Alg-MCC) composite was implanted to fill the dead space and support tissue regeneration. Postoperative healing was evaluated through clinical follow-up, computed tomography performed immediately and at 1 and 3 months postoperatively, as well as histological and immunohistochemical analyses. Imaging demonstrated progressive integration of the biomaterial, restoration of tissue density comparable to adjacent muscle, and absence of major complications. Histological evaluation demonstrated moderate inflammation and granulation tissue formation, containing numerous blood capillaries (neoangiogenesis), confirmed by CD31 and α-Smooth muscle actin (SMA) immunolabelling. Functional recovery was favorable, with preserved limb mobility and normal gait. These findings provide preliminary evidence of local biocompatibility and support the feasibility of incorporating the BGAuSP-Alg-MCC composite into a multimodal surgical management approach for extensive volumetric muscle loss in canine patients.

Early pregnancy detection in cattle is essential for improving reproductive efficiency, reducing days open, and enabling timely rebreeding of non-pregnant animals. Although several pregnancy diagnostic methods are available, none can reliably identify pregnancy before Day 21 post-insemination. Interferon tau (IFNT), the pregnancy recognition signal produced by the ruminant conceptus, initiates maternal responses through the induction of interferon-stimulated genes (ISGs). While this response is most pronounced in the endometrium, recent studies have shown that the cervix and vagina also respond to conceptus-derived IFNT, creating new possibilities for minimally invasive early pregnancy diagnosis. This review summarizes recent findings on the dynamics of ISG expression in the cervix and vagina, the detection of IFNT in cervico-vaginal secretions, and the diagnostic performance of these biomarkers. Available evidence indicates that cervical and vaginal tissues respond to conceptus-derived IFNT by upregulating the classical ISGs (ISG15, MX1, and MX2) as early as Days 14-16 post-insemination, coinciding with maternal recognition of pregnancy. Notably, cervical ISG15 expression on Day 16 demonstrates excellent diagnostic performance (AUC = 1.0). Furthermore, IFNT has been detected in cervical secretions on Days 16-19 with reported sensitivities of 94.5% in dairy cows and 96.6-100% in beef cows. The review also discusses the possible routes by which IFNT reaches the cervix and vagina and highlights the potential of IFNT-based lateral-flow assays for on-farm pregnancy diagnosis. Overall, the available evidence supports cervico-vaginal ISGs and IFNT as biologically grounded, minimally invasive biomarkers for the development of field-applicable early pregnancy diagnostic kits for dairy and beef cattle.

A 5-year-old jenny presented with dullness, depression, inappetence, stiffness of gait, and tachycardia (58 beats/min). Biochemical analysis revealed azotemia, hyperbilirubinemia, increased serum level of triglycerides (TG) and increased serum activities of aspartate aminotransferase (AST), creatinine phosphokinase (CPK), and γ-glutamyl transferase (GGT). Urinalysis showed ketonuria, bilirubinuria, proteinuria, and hyposthenuria. Ultrasonographic examination revealed loss of corticomedullary differentiation and slightly increased liver echogenicity. To the best of the author's knowledge, the present report is the first to detail the diagnostic findings, clinicopathological changes, therapeutic protocol, and clinical outcome of acute kidney injury and associated hyperlipidemia following exertional rhabdomyolysis in a jenny.

Malignant catarrhal fever (MCF) is a severe lymphoproliferative disease of domestic and wild ruminants caused by macaviruses, mainly ovine gammaherpesvirus 2 (OvHV-2) and alcelaphine gammaherpesvirus 1 (AlHV-1). Although relevant in sub-Saharan Africa, molecular epidemiological data remain limited, particularly in Namibia. This study investigated the occurrence and diversity of MCF viruses in Namibian livestock and contextualised local strains globally.Fifteen samples from cattle and sheep suspected of MCF were collected across five Namibian regions in 2023. Viral DNA was screened by duplex nested PCR, followed by amplification and sequencing of selected OvHV-2 (ORF50, ORF75, Ov9.5) and AlHV-1 (ORF50, A9.5) genomic regions. Phylogenetic and genetic distance analyses assessed viral diversity.OvHV-2 was the most commonly reported, being detected in 13/15 samples from both symptomatic cattle and asymptomatic sheep, whereas AlHV-1 was identified in two bovine cases. OvHV-2 sequences showed limited diversity and clustered into two main clades, without clear geographic structuring and with intermixing between host species. Several Namibian strains were closely related to sequences from Africa, Europe, Asia, and North America, depending on the genomic region analysed. AlHV-1 sequences were related to strains from southern and eastern Africa.These findings support widespread OvHV-2 circulation and indicate sheep as a likely infection source for cattle. The lack of spatial structure and presence of multiple clades suggest complex transmission dynamics, potentially influenced by animal movements, although the role of livestock trade and wildlife-mediated transmission remains unclear. Enhanced molecular surveillance, biosecurity, and control of animal translocations are needed to mitigate MCF impact in the region.

Vector-borne pathogens (VBPs) of medical and veterinary concern have mostly been studied in urban dog populations, while animals in rural settings, with limited access to ectoparasite prevention, have received less attention. In this study, we assessed canine exposure to VBPs in a rural Brazilian landscape. From August to October 2024, we collected 155 blood samples from dogs domiciled in a rural area of the municipality of Garanhuns, Pernambuco, Northeastern Brazil. Samples were assessed using an immunofluorescence antibody test (IFAT) to detect antibodies against Trypanosoma cruzi and using a rapid immunochromatographic test combined with an ELISA to detect Leishmania infantum antibodies. In addition, each sample was tested using a qualitative ELISA to detect D. immitis antigens and antibodies against Anaplasma phagocytophilum/Anaplasma platys, Ehrlichia canis/Ehrlichia ewingii, and Borrelia burgdorferi sensu lato. A total of 56.1% (87/155) of samples were reactive to at least one pathogen. Specifically, in 5.1% (8/155) were detected anti-T. cruzi antibodies, and in four of these samples (2.5%, 4/155) were co-detected antibodies anti-L. infantum. Antibodies against Ehrlichia spp. were observed in 50.9% (79/155), Anaplasma spp. in 8.3% (13/155), and B. burgdorferi in 0.6% (1/155). Antigens for D. immitis were detected in 1.2% (2/155). No statistical associations were observed on variables (sex, mobility, overnight stay and presence of ectoparasites) and pathogen seropositivity. In conclusion, dogs from rural areas are exposed to VBPs, which requires the use of ectoparasiticide preventive compounds to avoid ectoparasite infestation and reduce the risk of pathogen exposure.

The oral cavity provides a suitable environment for various microorganisms, including protozoa such as trichomonads, which may be associated with periodontal disease development. Humans are considered the typical hosts of Trichomonas tenax but it has also been found in dogs, cats, and other animals, which makes it a factor with a significant zoonotic potential. Apart from T. tenax, domestic carnivores may be infected with such species as T. canistomae, T. felistomae and T. brixi. The aim of this study was to determine the prevalence, species composition, and risk factors of trichomonad colonization of the oral cavity in dogs and cats from Poland. Oral swabs from 238 household animals (151 dogs and 87 cats) were examined using cultivation methods. Amplification of ITS1- 5.8 S rRNA-ITS2 fragment was performed for microscopic-positive samples and PCR products were further sequenced. Trichomonads were identified in 26 out of 151 (17.22%) canine and 5 out of 87 (5.75%) feline samples. Higher frequency was observed in older dogs and dogs with intense dental calculus. T. brixi and T. tenax were the only species detected in both dogs and cats. The detection of zoonotic T. tenax in a substantial number of examined animals highlights the relevance of this finding also in the context of human health.