Vet. Sci., Volume 10, Issue 3 (March 2023) – 68 articles

Streptococcus suis serotype 2 (SS2) is a noteworthy zoonotic pathogen that has been responsible for large economic losses in pig production and a great threat to human health. Pentraxin 3 (PTX3) is an essential regulator of the innate immune response to bacterial pathogens; however, its role during SS2 infection is not fully understood. In this study, we found that the SS2 strain HA9801 induced a significant inflammatory response in the mouse air pouch model; this response was amplified by the treatment of exogenous PTX3 simultaneously in terms of the results of inflammatory cell recruitment and proinflammatory cytokine IL-6 production. In addition, PTX3 facilitated the phagocytosis of macrophage Ana-1 against SS2 strain HA9801. The supplementation of exogenous PTX3 significantly reduced the bacterial loads in a dose-dependent manner in lungs, livers and bloods of SS2-infected mice compared to the samples with HA9801 infection alone; this finding indicated that PTX3 may facilitate the bacterial clearance through enhancing the host inflammatory response during SS2 infection. Both PTX3 and SS2 capsular polysaccharide (CPS2) were required for the robust inflammatory response, implying that the host PTX3 protein and SS2 surface CPS2 modulate the host innate immune response in concert. All of these results suggested that PTX3 is a potential novel biological agent for the SS2 infection; however, the recommended dose of PTX3 must be evaluated strictly to avoid inducing an excessive inflammatory response that can cause serious tissue injury and animal death. Full article

Our research purpose was to study the effect of the inclusion of a combination of phytobiotics in the form of dry Fucus vesiculosus grits (FG) and a mineral adsorbent from the heat-treated mineral shungite (TMS) on milk productivity, nutrient digestibility, and biochemical parameters of the Suksun dairy cows. A total of 80 dry-hardy cows of the Suksun breed were divided into four groups (20 heads each), balanced primarily by breed, age, body weight, body condition score, and indicators of milk yield for the previous lactation. The selected cows were with an average live body weight of 512.0 ± 1.28 kg, BCS 3.0–3.5, and parities of 6250 kg milk. The control group (CON) were fed the basic ration only; the second (TMS), third (FG), and fourth (TMS + FG) groups were fed the basic ration provided by 50 g of the mineral adsorbent from heat-treated shungite, 100 g of Fucus grits (Fucus vesiculosus), 50 g of the mineral adsorbent from heat-treated shungite, and 100 g of dry grits from Fucus vesiculosus, respectively. The total protein content in milk was significantly higher in the group receiving Fucus vesiculosus by 0.05% and the group receiving a combination of mineral adsorbent and Fucus vesiculosus by 0.03%. The percentage of milk fat content recorded the highest significant value in (TMS) group when compared to the control and represented (4.37 vs. 3.95). The group of cows that received (TMS + FG) revealed a significant difference in the digestibility of both ether extract and crude fiber when compared to the control group and represented (54.74 vs. 51.71 and 60.68 vs. 55.15%), respectively. The cows supplemented with a mineral adsorbent or a combination of mineral adsorbent and Fucus vesiculosus revealed a significant difference in the digestibility of ether extract and crude fiber in the group receiving TMS + FG by 3.0% (p < 0.05) and 5.5% (p < 0.05), respectively. The intake of nitrogen with the diet increased in (FG) and (TMS + FG) groups by 11.3 g (p < 0.05) and 13.4 g (p < 0.05) of nitrogen. There was an increase (p < 0.05) in the concentration of rumen ammonia in the control group compared to the other groups. The glucose content of those cows that received FG and TMS + FG combination increased (p < 0.05) by 0.76 mmol/L and 0.90 mmol/l in relation to the control group. The globulin, albumin/globulin ratio, and the level of triglycerides revealed a significant difference between the different experimental groups. In brief, the inclusion of a combination of phytobiotics in the form of dry Fucus vesiculosus grits and a mineral adsorbent from the heat-treated mineral shungite in Suksun dairy cows’ diets improved milk composition, digestibility of nutrients, utilization of nitrogen, and did not cause deleterious effects on blood biochemical indicators. Full article

Background: Toxoplasma gondii is classified as intracellular protozoa and is one of the major zoonotic parasites. Most warm-blooded intermediate hosts, including humans, are commonly infected by this parasite. The epidemiology of T. gondii infection in Egyptian horses is currently poorly understood. Methods: 420 blood samples were randomly collected from horses raised in four governorates in Northern Egypt (110 each from Giza and Kafr El Sheikh, and 100 each from Qalyubia and Gharbia) to investigate the existence of antibodies against T. gondii using a commercial ELISA kit, and to ascertain the risk factors for the infection. Results: the antibodies for T. gondii were found in 16.2% (68/420) of the examined horses, with no significant differences among the four studied governorates. The highest prevalence rate was observed in Giza. The results revealed that sex, breed, age, and contact with domestic ruminants or cats were recognized as potential risk factors. The high prevalence rate was found in mixed breed horses (OR = 2.63, 95% CI: 0.95–7.26), mares (OR = 2.35, 95% CI: 1.31–4.19), and horses aged over 10 years (OR = 2.78, 95% CI: 1.30–3.44). Moreover, the likelihood of seropositivity for T. gondii infection was higher in horses raised in environments with cats (OR = 1.97, 95% CI: 1.13–3.44, p = 0.017) or domestic ruminants (OR = 2.16, 1.21–3.86, p = 0.010). This report confirms that horses in Northern Egypt are exposed to T. gondii and thus raises the possibility that people and other animals could contract the disease. Conclusions: routine examination and management of T. gondii infection in horses in these governorates is advised. Full article

Virulent Aeromonas hydrophila (vAh) is a major bacterial pathogen in the U.S. catfish industry and is responsible for large-scale losses within commercial ponds. Administering antibiotic feeds can effectively treat vAh infections, but it is imperative to discern new approaches and better understand the mechanics of infection for this bacterium. As such, the persistence of vAh in pond sediments was determined by conducting laboratory trials using sediment from four commercial catfish ponds. Twelve chambers contained sterilized sediment, vAh isolate ML-09-119, and 8 L of water maintained at 28 °C and were aerated daily. At 1, 2, 4, 6, and 8 days, and every 7th day post-inoculation for 28 days, 1 g of sediment was removed, and vAh colony forming units (CFU) were enumerated on ampicillin dextrin agar. Viable vAh colonies were present in all sediments at all sampling periods. The vAh growth curve peaked (1.33 ± 0.26 × 10⁹ CFU g⁻¹) at 96 h post-inoculation. The population plateaued between days 14 and 28. No correlations were found between CFU g⁻¹ and physiochemical sediment variables. This study validated the ability of vAh to persist within pond sediments in a laboratory setting. Further research on environmental factors influencing vAh survivability and population dynamics in ponds is needed. Full article

The macrophage CD163 surface glycoprotein is a member of the SRCR family class B, which has been identified as the key trigger in host–pathogen interactions, but its specific roles in sensing Glaesserella parasuis (G. parasuis) infection are largely unknown. Here, we investigated porcine CD163 in mediating the adhesion and immune response of G. parasuis using in vitro host–bacteria interaction models. CD163-overexpressing Chinese hamster ovary K1 cells (CHO-K1) showed obvious subcellular localization in the cytoplasm, especially in the cytomembrane. Although detection using scanning electron microscopy (SEM) confirmed the bacterial adhesion, there was no significant difference in the adhesion of G. parasuis to CHO-K1 cells between the presence and absence of CD163. In addition, similar results were observed in 3D4/21 cells. Meanwhile, bindings of G. parasuis to nine synthetic peptides, the bacterial binding motifs within SRCR domains of CD163, were weak based on a solid-phase adhesion assay and agglutination assay. Moreover, CD163 had no effect on the expression of G. parasuis-induced inflammatory cytokines (IL-6, INF-γ, IL-10, IL-4 and TGF-β) in CHO-K1 cells. In conclusion, these findings indicate that porcine CD163 plays a minor role in sensing G. parasuis infection. Full article

While human and animal leishmaniasis affect several millions of people worldwide, L. infantum is the species responsible for visceral leishmaniasis in Europe, Middle East, and America. Antileishmanial drugs present issues associated with drug toxicity and increasing parasite resistance. Therefore, the study of this parasite with a focus on new potential drug targets is extremely useful. Accordingly, we purified and characterized a transglutaminase (TGase) from L. infantum promastigotes. While Tgases are known to be involved in cell death and autophagy, it appears that these functions are very important for parasites’ virulence. For the first time, we showed a Ca²⁺- and GTP-dependent TGase in Leishmania corresponding to a 54 kDa protein, which was purified by two chromatographic steps: DEAE-Sepharose and Heparin-Sepharose. Using polyclonal antibodies against a 50-amino-acid conserved region of the catalytic core of human TGase 2, we revealed two other bands of 66 and 75 kDa. The 54 kDa band appears to be different from the previously reported TGase, which was shown to be Ca²⁺- independent. Future research should address the identification of the purified enzyme sequence and, subsequently, its cloning to more comprehensively investigate its pathophysiological function and possible differences from mammal enzymes. Full article

Acute diarrhea is a very frequent condition affecting dogs; nevertheless, little is known about what happens in the GI tract during such conditions. Proteomics allows the study of proteins present in a specific biologic substrate, and fecal proteomic investigations have been recently implemented to study GI diseases in dogs. In the present study, the fecal protein profiles of eight dogs suffering from acute uncomplicated diarrhea at the time of inclusion was investigated for the first time, and then the same patients were followed, replicating two further evaluations at two subsequent time points (after 2 and 14 days from the first presentation), with the aim of gaining possible new insights regarding the pathologic changes in the gastrointestinal environment during such conditions. Two-dimensional gel electrophoresis (2-DE) was performed, followed by mass spectrometry. Nine spots, corresponding to four (groups of) proteins (i.e., albumin, alkaline phosphatase, chymotrypsin-C-like, and some immunoglobulins), showed significant differences at two or more of the three time points investigated, almost all behaving similarly and decreasing at T1 (2 days after the onset of the condition) and significantly increasing at T2 (14 days after the onset), mainly evidencing a reaction of the organism. Further studies including a greater number of patients and possibly different techniques are needed to confirm the present findings. Full article

Cats urgently visit emergency hospitals due to respiratory distress, and the chief cause is cardiogenic pulmonary edema (CPE). Although cats with CPE were frequently encountered in clinics, the prognostic factors were poorly reported. The objective of this retrospective study was to investigate the association of physical examination and venous blood gas parameters with the survival of cats with CPE in an emergency hospital. Thirty-six cats with CPE were ultimately included in the present study, and eight of them died within 12 h after their presentation to our hospital. Statistical analyses of clinical parameters between cats that died within 12 h and those that survived for 12 h were conducted using Mann–Whitney U test with Bonferroni correction. Cats that died within 12 h had significantly lower rectal temperatures and higher PvCO₂ than those that did not die within 12 h. Moreover, hypotension and vasoconstrictor use were related to death within 12 h of presentation and higher PvCO₂. These findings indicated the prognostic utility of body temperature and PvCO₂, and the association between hypercapnia and the severity of CPE or hypotension. A large number of prospective studies should be performed to validate these results. Full article

The objectives of this study were (1) to investigate the distribution of large (≥10 mm) follicle numbers during the estrous cycle and (2) to compare the timing of the estrus expression period after the ovarian examination between cows with one large follicle (1F) and two or more large follicles (2F) with functional corpus luteum (CL) at the ovarian examination in lactating Holstein dairy cows. In experiment 1, we performed 393 ovarian examinations by ultrasonography, addressed the existence of CL (≥20 mm) and large follicle numbers, and classified cows into 1F (n = 229) and 2F (n = 164) groups. The 1F appearance rates were beyond 75% each day during 3 to 12 d after estrus. However, 2F appearance rates were beyond 75% each day during 15 to 24 d after estrus. In experiment 2, we performed 302 ovarian examinations by ultrasonography and classified cows into the 1F (n = 168) and 2F (n = 134) groups. Estrus detection was performed for 24 d after the ovarian examination in each cow. In the 2F group, 75% of estrus occurred within 9 d of the ovarian examination. However, 75% of estrus occurred 10 d after the ovarian examination in 1F. Days from the ovarian examination to estrus were significantly shorter in the 2F (6.0 d; median, 7.2 ± 4.0 d; mean ± SD) than in the 1F (13 d, 12.4 ± 4.3 d) group. In conclusion, focusing on ≥10 mm follicle numbers with CL could be useful for predicting the estrus expression period. Full article

Wild animals harbor pathogens that can be infectious agents for humans, including parasites. This study aimed to identify gastrointestinal parasites and assess their prevalence and the potential risk for humans associated with consuming these animals. The research was conducted from August to December 2019. Parasitological analyses were carried out on the feces and intestines of 113 wild animals, including antelopes (24), duikers (58), porcupines (18), small monkeys (Cercopithecus) (8), nandinia (2), pangolin (1), genet (1), and a crocodile (1), from the Zadié Department in the province of Ogooué-Ivindo in the northeast of Gabon. The results revealed 15 taxa of gastrointestinal parasites, including nine nematodes: Strongylids (61/113), Strongyloides spp. (21/113), Ascaris spp. (21/113), Trichuris spp. (39/113), Capillaria spp. (9/113), Protostrongylus spp. (5/113), Enterobius spp. (8/113), Toxocara spp. (7/113) and Mammomonogamus spp. (5/113); three species of protozoa, namely Balantidium spp. (12/113), Eimeria spp. (17/113), and Entamoeba spp. (9/113); two species of trematodes, namely Fasciola spp. (18/113) and Paramphistomum spp. (21/113); and cestode species, Taenia spp. (1/113). The prevalence of gastrointestinal parasitism in these animals was 85.84% (97/113). In addition, among these parasitic taxa, some are potential pathogens for humans, such as Ascaris spp., Balantidium spp., Entamoeba spp., and Taenia spp. The consumption of games, particularly offal, infested by these parasites, could threaten human health. Full article

Pulmonary disease is often associated with feedlot cattle mortality, and the most common syndromes include bronchopneumonia, acute interstitial pneumonia, and bronchopneumonia with an interstitial pneumonia. The study objective was to utilize gross necropsy and histopathology to determine the frequency of pulmonary lesions from three major syndromes and agreement between gross and histopathological diagnosis. A cross sectional, observational study was performed at six U.S. feedyards using a full systematic necropsy to assess mortalities during summer 2022. A subset of mortalities had four lung samples submitted for histopathological diagnosis. Gross necropsy was performed on 417 mortalities, 402 received a gross diagnosis and 189 had a histopathological diagnosis. Descriptive statistics were used to evaluate pulmonary diagnosis frequency based on method (gross/histopathology), and generalized linear mixed models were used to evaluate agreement between histopathological and gross diagnoses. Using gross diagnosis, bronchopneumonia represented 36.6% of cases with acute interstitial pneumonia and bronchopneumonia with an interstitial pneumonia representing 10.0% and 35.8%, respectively. Results identified bronchopneumonia with an interstitial pneumonia as a frequent syndrome which has only been recently reported. Histopathological diagnosis had similar findings; bronchopneumonia represented 32.3% of cases, with acute interstitial pneumonia and bronchopneumonia with an interstitial pneumonia representing 12.2% and 36.0%, respectively. Histopathological diagnosis tended (p-VALUE = 0.06) to be associated with gross diagnosis. Pulmonary disease was common and both diagnostic modalities illustrated three primary syndromes: bronchopneumonia, acute interstitial pneumonia, and bronchopneumonia with an interstitial pneumonia with similar frequencies. Improved understanding of pulmonary pathology can be valuable for evaluating and adjusting therapeutic interventions. Full article

The objective of our study was to survey Babesia infection rates by PCR and tick species on stray dogs to correlate the distribution of Babesia with the distribution of ticks infesting dogs in Taiwan. Three hundred eighty-eight blood samples and 3037 ticks were collected from 388 roaming, and free-ranging owned dogs at residential sites in Taiwan between January 2015 and December 2017. The prevalence of B. gibsoni and B. vogeli was 15.7% (61/388) and 9.5% (37/388), respectively. Most positive B. gibsoni dogs were found in the northern part of the country 56/61 (91.8%), whereas a few were found in the middle 5/61 (8.2%). Babesia vogeli infection rates were 10%, 3.6%, and 18.2% in the northern, central, and southern regions, respectively. Five species of ticks were found: Rhipicephalus sanguineus (throughout Taiwan), Rhipicephalus haemaphysaloides (in the north), Haemaphysalis hystricis (in the north and middle of Taiwan), and Amblyomma testidunarium and Ixodes ovatus (both in the north). None of the dogs in the south were infected with B gibsoni, which correlated with the absence of H. hystricis, a tick recently identified as the local vector for B gibsoni. Babesia vogeli was more equally distributed, coinciding with R. sanguineus, a tick that is present throughout Taiwan. Anaemia was detected in 86.9% of infected dogs; among these dogs, approximately 19.7% showed severe anaemia (HCT < 20). These findings provide useful advice for owners regarding outdoor activities with their dogs and local veterinarians with a regional differential diagnosis of babesiosis in Taiwan. Full article

This study aimed to determine how milk composition, milk microbiota, and blood metabolites may change during the lactation period in Jersey cows. Milk and jugular blood samples were collected from eight healthy cows every other month from the beginning to the end of their lactation period. Samples of airborne dust were also collected to determine whether the cowshed microbiota could affect milk microbiota. Milk yield peaked in the first two months and gradually decreased as the lactation period progressed. Milk fat, protein, and solids-not-fat contents were low in the first month, and then increased during the middle and late lactation periods. In the first month, plasma non-esterified fatty acids (NEFA), haptoglobin (Hp), and aspartate transaminase (AST) levels were elevated, and high abundances of Burkholderiaceae and Oxalobacteraceae were observed in milk and airborne dust microbiota. The finding that contamination of the environmental microbiota in milk was coupled with elevated plasma NEFA, Hp, and AST levels indicated that impaired metabolic function during the early lactation period may increase the invasion of opportunistic bacteria. This study can affirm the importance of feeding and cowshed management and should provide a helpful addition to improving Jersey cow farming. Full article

The transition dairy cows are challenged by various stresses such as decreased dry matter intake, liver dysfunction, increased inflammation, and oxidative stress, particularly in subtropical regions. These might increase the requirement for vitamin E and trace elements. To examine whether supplementation of vitamin E, selenium or copper, zinc, and manganese complex would help transition dairy cows to achieve greater reproduction performance by overcoming the immune function and postpartum disorders in subtropical Taiwan. A total of 24 Holstein Friesian dairy cows were enrolled in this study and divided into three groups (n = 8 cows/group): treatment 1 supplemented with organic selenium and vitamin E (SeE), treatment 2 supplemented with organic copper, zinc, and manganese complex (CZM) and control (CON). The results showed SeE supplementation improved immune function, reproductive performance, and milk yield, but not negative energy balance status. Supplementation of CZM improved milk yield and energy regulation through antioxidative capacity and immune function, but had no influence on reproductive performance. Full article

To analyze the intervention mechanism of polysaccharides from charred Angelica sinensis (CASP) on the liver injury caused by Ceftiofur sodium (CS) and lipopolysaccharide (LPS) from the perspective of the intestine. Ninety-four one-day-old laying chickens underwent free feeding and drinking water for three days. Then, fourteen laying chickens were randomly selected as the control group, and sixteen laying chickens were selected as the model group. Sixteen laying chickens in the rest were randomly selected as the intervention group of CASP. Chickens in the intervention group were given CASP by the oral administration (0.25 g/kg/d) for 10 days, the control and model groups were given the same amount of physiological saline. During the 8th and 10th days, laying chickens in the model and CASP intervention group were subcutaneously injected with CS at the neck. In contrast, those in the control group were subcutaneously injected with the same amount of normal saline simultaneously. Except for the control group, the layer chickens in the model and CASP intervention groups were injected with LPS after CS injection on the 10th day of the experiment. In contrast, those in the control group were injected with the same amount of normal saline at the same time. 48 h after the experiment, the liver samples of each group were collected, and the liver injury was analyzed by hematoxylin-eosin (HE) staining and transmission electron microscopy. And the cecum contents of six-layer chickens in each group were collected, and the intervention mechanism of CASP on the liver injury from the perspective of the intestine was analyzed by the 16S rDNA amplicon sequencing technology and the short-chain fatty acids (SCFAs) detection of cecal contents based on Gas Chromatography-Mass Spectrometry (GC-MS), and their association analysis was carried out. The results showed that the structure of chicken liver in the normal control group was normal, while that in the model group was damaged. The structure of chicken liver in the CASP intervention group was similar to the normal control group. The intestinal floras in the model group were maladjusted compared to the normal control group. After the intervention of CASP, the diversity, and richness of chicken intestinal floras changed significantly. It was speculated that the intervention mechanism of CASP on the chicken liver injury might be related to the abundance and proportion of Bacteroidetes and Firmicutes. Compared with the model group, the indexes of ace, chao1, observed species, and PD whole tree of chicken cecum floras in the intervention group of CASP were significantly increased (p < 0.05). The contents of acetic acid, butyric acid, and total SCFAs in the intervention group of CASP were significantly lower than those in the model group (p < 0.05), and the contents of propionic acid and valeric acid in the intervention group of CASP were significantly lower than those in the model group (p < 0.05) and normal control group (p < 0.05). The correlation analysis showed that the changes in the intestinal floras were correlated with the changes in SCFAs in the cecum. It is confirmed that the liver-protecting effect of CASP is indeed related to the changes in the intestinal floras and SCFAs content in the cecum, which provides a basis for screening liver-protecting alternative antibiotics products for poultry. Full article

Avian orthoavulavirus-1 (AOAV-1) is the causative agent of Newcastle disease in poultry. This highly infectious disease causes large economic losses annually and worldwide. AOAV-1 does not only infect poultry, but it has a very broad host range and has been detected in over 230 bird species to date. A distinct group of viral strains within AOAV-1 are pigeon-adapted strains, also named pigeon paramyxovirus-1 (PPMV-1). AOAV-1 is transmitted through the feces of infected birds and secretions from the nasal and oral cavities and eyes. It is worth mentioning that wild birds can transmit the virus to captive birds, especially feral pigeons to poultry. Therefore, early and sensitive detection of this virus—including the monitoring of pigeons—is of utmost importance. A variety of molecular methods for the detection of AOAV-1 already exist, but the detection of the F gene cleavage site of currently circulating PPMV-1 strains has not proven to be particularly sensitive or suitable. As presented here, by modifying the primers and probe of an already established real-time reverse-transcription PCR, the sensitivity could be increased, allowing for a more reliable detection of the AOAV-1 F gene cleavage site. Furthermore, it becomes clear how important it is to constantly monitor and, if necessary, adapt existing diagnostic procedures. Full article

Transcutaneous abdominal ultrasonography using alcohol saturation is used in the diagnostic workup of a variety of conditions in horses. The duration of the examination and the amount of alcohol used in each case may vary depending on several factors. The aim of this study is to describe the breath alcohol test results obtained by veterinarians performing abdominal ultrasound on horses. Six volunteers were enrolled, after written consent, and a Standardbred mare was used for the whole study protocol. Each operator performed a total of 6 ultrasounds by pouring the ethanol solution from a jar or by spray application, for a duration of 10, 30, and 60 min. An infrared breath alcohol analyzer was used immediately after completing the ultrasonography and at 5-min intervals until a negative result was obtained. Positive results were obtained for 0–60 min after the procedure. A statistically significant difference was found between the groups that used more than 1000 mL, 300–1000 mL, and less than 300 mL of ethanol. No significant differences were observed between the type of ethanol administration and the time of exposure. Based on this study, equine vets who perform ultrasound on horses can test positive at the breath alcohol test for up to 60 min following ethanol exposure. Full article

OmpH is among the most important virulence factors of Pasteurella multocida, which mediates septicemia in yaks (Bos grunniens I) after infection with the bacteria. In the present study, yaks were infected with wild-type (WT) (P0910) and OmpH-deficient (ΔOmpH) P. multocida strains. The mutant strain was generated through the reverse genetic operation system of pathogens and proteomics technology. The live-cell bacterial count and clinical manifestations of P. multocida infection in Qinghai yak tissues (thymus, lung, spleen, lymph node, liver, kidney, and heart) were analyzed. The expression of differential proteins in the yak spleen under different treatments was analyzed using the marker-free method. We found that compared with the mutant strain, the titer of wild-type strains was significantly higher in tissues. Additionally, compared with other organs, the bacteria titer was significantly higher in the spleen. Compared with the WT p0910 strain, the mutant strain generated milder pathological changes in the tissues of yak. Proteomics analysis revealed that 57 of the 773 proteins expressed in P. multocida were significantly differentially expressed between the ΔOmpH and P0910 groups. Of the 57, 14 were over-expressed, whereas 43 were under-expressed. The differentially expressed proteins in the ΔompH group regulated the ABC transporter (ATP-powered translocation of many substrates across membranes) system, the two-component system, RNA degradation, RNA transcription, glycolysis/gluconeogenesis, biosynthesis of ubiquinone and other terpenoid-quinones, oxidative phosphorylation (citrate cycle) as well as fructose and mannose metabolism. The relationship among 54 significantly regulated proteins was analyzed using STRING. We found that WT P0910 and ΔOmpH of P. multocida infection activated the expression of ropE, HSPBP1, FERH, ATP10A, ABCA13, RRP7A, IL-10, IFN-γ, IL-17A, EGFR, and dnaJ. Overall, deletion of the OmpH gene weakened the virulence but maintained the immunogenicity of P. multocida in yak. The findings of this study provide a strong foundation for the pathogenesis of P. multocida and the management of related septicemia in yaks. Full article

Point-of-care diagnostic technologies are becoming more widely available for production species. Here, we describe the application of reverse transcription loop-mediated isothermal amplification (RT-LAMP) to detect the matrix (M) gene of influenza A virus in swine (IAV-S). M-specific LAMP primers were designed based on M gene sequences from IAV-S isolated in the USA between 2017 and 2020. The LAMP assay was incubated at 65 °C for 30 min, with the fluorescent signal read every 20 s. The assay’s limit of detection (LOD) was 20 M gene copies for direct LAMP of the matrix gene standard, and 100 M gene copies when using spiked extraction kits. The LOD was 1000 M genes when using cell culture samples. Detection in clinical samples showed a sensitivity of 94.3% and a specificity of 94.9%. These results show that the influenza M gene RT-LAMP assay can detect the presence of IAV in research laboratory conditions. With the appropriate fluorescent reader and heat block, the assay could be quickly validated as a low-cost, rapid, IAV-S screening tool for use on farms or in clinical diagnostic labs. Full article

D-532 fertilization solution is generally used to replace the water or ovarian fluid during artificial reproductive practices in salmonids due to its ability to boost sperm motility and increase fertilization rates compared with natural activation media. However, the maintenance of ovarian fluid in a reproductive microenvironment gives it the advantage of protecting the eggs from potential harmful factors from the external environment and simplifying the field operations related to its removal when D-532 is used alone. In light of this, the aim of the present study was to investigate in vitro, for the first time, the effect of ovarian fluid (OF 100%) on post-thaw sperm swimming performance of Mediterranean trout, comparing it with D-532 and a mixed solution of 50% D-532 and 50% ovarian fluid (OF 50%). The percentage of motile spermatozoa and movement duration was significantly increased in OF 100% and OF 50% compared with D-532. Sperm velocity was higher in D-532, but significant differences were recorded only with OF 100%. In conclusion, these results suggest that the presence of ovarian fluid alone or in combination with D-532 in an artificial microenvironment of reproduction represents a key factor in potentially increasing fertilization success when the frozen semen of Mediterranean brown trout is used. Full article

Galectins are proteins that bind to glycans in targeted cells and function in cell-to-cell signaling throughout the body. Galectins have been found to be involved in various reproductive processes, including placental dysfunction, but this has not been investigated in the horse. Therefore, the objective of this study was to assess alterations in galectin expression of the abnormal placenta in pregnant mares. Next-generation RNA sequencing was performed on the postpartum chorioallantois of two placental pathologies following clinical cases of ascending placentitis (n = 7) and focal mucoid placentitis (n = 4), while chorioallantois from healthy postpartum pregnancies (n = 8; 4 control samples per disease group) served as the control. When evaluating ascending placentitis, both galectin-1 (p < 0.001) and galectin-3BP (p = 0.05) increased in the postpartum chorioallantois associated with disease, while galectin-8 (p < 0.0001) and galectin-12 (p < 0.01) decreased in the diseased chorioallantois in comparison with those in the control. In mares with focal mucoid placentitis, numerous galectins increased in the diseased chorioallantois, and this included galectin-1 (p < 0.01), galectin-3BP (p = 0.03), galectin-9 (p = 0.02), and galectin-12 (p = 0.04), in addition to a trend toward increases in galectin-3 (p = 0.08) and galectin-13 (p = 0.09). In contrast, galectin-8 expression decreased (p = 0.04) in the diseased chorioallantois in comparison with that of the controls. In conclusion, galectins alter in abnormal placentae with variations observed among two forms of placental pathologies. These cytokine-like proteins may further our understanding of placental pathophysiology and warrant attention as potential markers of placental inflammation and dysfunction in the horse. Full article

The tooth is made up of three mineralized tissues, enamel, dentin, and cementum, which surround a non-mineralized tissue called the dental pulp. Micro-computed tomography (mCT) is an imaging technology based on X-rays that allows non-invasive visualization of objects at a microscopic scale, according to their radiopacity and in three dimensions (3D). Likewise, it allows the subsequent execution of morphological and quantitative analysis of the objects, such as, for example, the determination of the relative mineral density (MD). The present work aimed to describe the MD of feline teeth using mCT. The studied sample consisted of four European Shorthair cats, from which nine canine teeth were extracted per medical indication. These teeth were evaluated through dental radiography before and after their extraction. Using mCT and the CTAn software, the values of the relative mineral density of the root of each tooth and of specific segments corresponding to the coronal, middle, and apical thirds of the root were determined. Mean MD of root tissues was 1.374 ± 0040 g·cm⁻³, and of hard root, tissues was 1.402 ± 0.035 g·cm⁻³. Through mCT, it was possible to determine the mean MD values of feline canine teeth. The study of MD could become an ancillary method for the diagnosis and characterization of dental pathology. Full article

Otitis media can be a consequence of chronic otitis externa and could represent a perpetuating factor. While the microbiota of the EEC in healthy dogs and in the presence of otitis externa has been described, only sparse information is available concerning the normal microbiota of the middle ear. The objective was to compare the tympanic bulla (TB) with the external ear canal (EEC) microbiota in healthy dogs. Six healthy experimental Beagle dogs were selected based on the absence of otitis externa, negative cytology and bacterial culture from the TB. Samples from the EEC and TB were collected directly after death using a total ear canal ablation and lateral bulla osteotomy. The hypervariable segment V1–V3 of the 16S rDNA was amplified and sequenced with a MiSeq Illumina. The sequences were analyzed by the Mothur software using the SILVA database. No significant differences between the EEC and TB microbiota for the Chao1 richness index (p = 0.6544), the Simpson evenness index (p = 0.4328) and the reciprocal Simpson alpha diversity (p = 0.4313) were noted (Kruskal-Wallis test). A significant difference (p = 0.009) for the Chao1 richness index between the right and left EEC was observed. The microbiota profile was similar in the EEC and the TB of the Beagles. Full article

Endometritis is one of the most important causes of infertility in dairy cows, resulting in high economic losses in the dairy industry. Though the presence of a commensal uterine microbiota is now well established, the complex role of these bacteria in genital health, fertility, and susceptibility to uterine diseases remains unclear. In this study, we explore the endometrial microbiota through 16S rRNA gene profiling from cytobrush samples taken ex vivo from healthy, pregnant, and endometritis cows. There were no significant differences between healthy and pregnant cows, whose uterine microbiota were dominated by Streptococcus, Pseudomonas, Fusobacterium, Lactococcus and Bacteroides. Compared to pregnant and clinically healthy cows, the uterine bacterial community of endometritis cows was significantly decreased in species diversity (p < 0.05), reflecting uneven community composition in different patterns with either dominance of Escherichia-Shigella, Histophilus, Bacteroides and Porphyromonas or Actinobacteria. Full article

Activation of the AMP-activated protein kinase (AMPK) has been demonstrated to be beneficial for boar sperm quality and functionality, while the underlying mechanism of AMPK activation of boar spermatozoa remains obscure. This study aimed to explore the effect of antioxidants and oxidants in boar spermatozoa and their surrounding fluid (SF) on the activation of AMPK during the liquid storage. Ejaculates from Duroc boars, routinely used for semen production, were collected and diluted to a final concentration of 25 × 10⁶/mL. In experiment 1, twenty-five semen samples from eighteen boars were stored at 17 °C for 7 days. In experiment 2, three pooled semen samples created from nine ejaculates of nine boars were used, and each sample was treated with 0, 0.1, 0.2, and 0.4 μM/L H₂O₂ and stored at 17 °C for 3 h. Sperm quality and functionality, antioxidants and oxidants in boar spermatozoa and SF, the intracellular AMP/ATP ratio, and the expression levels of the phosphorylated AMPK (Thr172) were determined. Sperm quality significantly decreased with storage time in terms of viability (p < 0.05). Antioxidant and oxidant levels were markedly affected with storage time, with a decline in the SF total antioxidant capacity (TAC) (p < 0.05), SF malondialdehyde (MDA) (p < 0.05), and the sperm’s total oxidant status (TOS), as well as a fluctuation in sperm superoxidase dismutase-like (SOD-like) activity (p < 0.05). The intracellular AMP/ATP ratio increased (p < 0.05) on day 4 and subsequently decreased to its lowest value on days 6 and 7 (p < 0.05). The phosphorylated AMPK levels increased from day 2 to day 7 (p < 0.05). Correlation analyses indicate that sperm quality during liquid storage was correlated to antioxidants and oxidants in spermatozoa and SF (p < 0.05), which were correlated to the phosphorylation of sperm AMPK (p < 0.05). Treatment with H₂O₂ induced damages in sperm quality (p < 0.05), a decline in antioxidant levels (SF TAC, p < 0.05; sperm SOD-like activity, p < 0.01), an increase in oxidant levels (SF MDA, p < 0.05; intracellular ROS production, p < 0.05), a higher AMP/ATP ratio (p < 0.05), and phosphorylated AMPK levels (p < 0.05) in comparison with the control. The results suggest that antioxidants and oxidants in boar spermatozoa and SF are involved in AMPK activation during liquid storage. Full article

American foulbrood is caused by the spore-forming Paenibacillus larvae. Although the disease effects honey bee larvae, it threatens the entire colony. Clinical signs of the disease are seen at a very late stage of the disease and bee colonies are often beyond saving. Therefore, through active monitoring based on screening, an infection can be detected early and bee colonies can be protected with hygiene measures. As a result, the pressure to spread in an area remains low. The cultural and molecular biological detection of P. larvae is usually preceded by spore germination before detection. In this study, we compared the results of two methods, the culture detection and RT-PCR detection of DNA directly isolated from spores. Samples of honey and cells with honey surrounding the brood were used in a five-year voluntary monitoring program in a western part of Lower Austria. DNA-extraction from spores to speed up detection involved one chemical and two enzymatic steps before mechanical bashing-beat separation and additional lysis. The results are comparable to culture-based methods, but with a large time advantage. Within the voluntary monitoring program, the proportion of bee colonies without the detection of P. larvae was high (2018: 91.9%, 2019: 72.09%, 2020: 74.6%, 2021: 81.35%, 2022: 84.5%), and in most P. larvae-positive bee colonies, only a very low spore content was detected. Nevertheless, two bee colonies in one apiary with clinical signs of disease had to be eradicated. Full article

The purpose of this study was to determine the level of application and effectiveness of the use of vegetable feed additives from complex phytobiotic feed additives (CPFA) in the diets of broiler chickens, as well as their effects on growth indicators, carcass characters and blood profile. A total of 258 Ross 308 chicks were divided into six dietary regimens, including: a basal diet without additives as a first control group (CON); the second group received a basal diet supplemented with 200 g/t in the starter phase and 100 g/t in the grower and finisher phase; the third group—400 g/t and 200 g/t; the fourth group—600 g/t and 300 g/t; the fifth group—800 g/t and 400 g/t; and the sixth group—1000 g/t and 500 g/t of a complex phytobiotic supplement based on tannins, respectively. The CPFA contains the following: tannins 36.8–55.2%, eugenol 0.4–0.6%, cinnamon aldehyde 0.8–1.2%, zinc–methionine 1.6–2.4%, calcium butyrate 8–12%, silicon dioxide 1.2–1.8% and dextrose up to 100%. The maximum introduction of phytobiotics (1000 g/t) at 7 days of age leads to a decrease in the live weight of broilers which reduced by 8.27% (p < 0.05) compared to the minimum level of phytobiotics (200 g/t). From 15–21 days, the live weight was significant between the supplemented and control groups and represented 396.21, 384.81 and 384.16 vs. 316.91 g for the CPFA 4, CPFA 5, CPFA 1 and control group, respectively. Furthermore, the same trend was recorded in the average daily gain during the periods between 15–21 and 22–28 days of the experiment. Feeding CPFA had a positive effect on the carcass indicators, except for the feeding of CPFA 3 in the amount of 600 g/t in the starter phase and 300 g/t in the grower and finish phases, which recorded the lowest weight in relation to the CPFA 1 and 2 groups and represented 1309.58 vs. 1460.06 and 1456.52 g, respectively, and the difference was significant. The inclusion of CPFA in poultry diets contributed to an increase in lung mass in the experimental groups relative to the control group, except for the CPFA 5 group which represented the lowest weight of lung mass (6.51 g) and the differences were significant between the CPFA 2 and CPFA 3 and the control groups. The highest concentration of leukocytes was observed during the experiment period in the group of poultry receiving phytobiotics (CPFA 3), which significantly exceeded the control group by 2.37 × 109/L. A significant decrease in the level of cholesterol was recorded in the CPFA groups when compared to the control group and represented 2.83 vs. 3.55 mmol/L, respectively. Consequently, the introduction of vegetable feed additives from complex phytobiotic feed additives (CPFA) in the diets of Ross 308 chicks had a positive effect on the growth production, the carcass yield, the mass of the pectoral muscles and the mass of the lungs. Moreover, it did not cause a harmful effect on the biochemical parameters of the blood. Full article

Bovine respiratory disease (BRD) remains the leading disease within the U.S. beef cattle industry. Marketing decisions made prior to backgrounding may shift BRD incidence into a different phase of production, and the importance of host gene expression on BRD incidence as it relates to marketing strategy is poorly understood. Our objective was to compare the influence of marketing on host transcriptomes measured on arrival at a backgrounding facility on the subsequent probability of being treated for BRD during a 45-day backgrounding phase. This study, through RNA-Seq analysis of blood samples collected on arrival, evaluated gene expression differences between cattle which experienced a commercial auction setting (AUCTION) versus cattle directly shipped to backgrounding from the cow–calf phase (DIRECT); further analyses were conducted to determine differentially expressed genes (DEGs) between cattle which remained clinically healthy during backgrounding (HEALTHY) versus those that required treatment for clinical BRD within 45 days of arrival (BRD). A profound difference in DEGs (n = 2961) was identified between AUCTION cattle compared to DIRECT cattle, regardless of BRD development; these DEGs encoded for proteins involved in antiviral defense (increased in AUCTION), cell growth regulation (decreased in AUCTION), and inflammatory mediation (decreased in AUCTION). Nine and four DEGs were identified between BRD and HEALTHY cohorts in the AUCTION and DIRECT groups, respectively; DEGs between disease cohorts in the AUCTION group encoded for proteins involved in collagen synthesis and platelet aggregation (increased in HEALTHY). Our work demonstrates the clear influence marketing has on host expression and identified genes and mechanisms which may predict BRD risk. Full article