Intestine Of Cattle Research Articles

Assessment of Potential Probiotic Yeasts Isolated from The Small Intestines of Cattle in Thailand

Y easts are widely studied and considered good candidates for probiotics due to their numerous benefits. This study aimed to investigate potential probiotic attributes of yeasts isolated from the small intestines of cattle in Thailand. Growth at body temperature, resistance to low pH, auto-aggregation capability, hydrophobicity, growth under gastrointestinal tract (GIT) conditions, safety profiles, antimicrobial activity, and extracellular enzyme production of yeast strains were evaluated in vitro. The results of sequential screening indicated that 829 out of 1,025 yeast strains could grow at both 37 and 39 °C, 682 strains could resist acidity at pH 2 for 24 h, 252 strains revealed greater than 83% auto-aggregation capability, and 97 strains showed higher than 90% hydrophobicity. Sixty strains grew well under simulated GIT conditions in the presence of pepsin, pancreatin, and bile salts. Thirteen strains of Diutina rugosa showed negative results for hemolytic, gelatinase, and phospholipase activities, while seven strains of Pichia kudriavzevii also showed negative results for the above activities, except for phospholipase. Neither D. rugosa nor P. kudriavzevii inhibited Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. Additionally, 12 strains of D. rugosa and four strains of P. kudriavzevii produced phytase and cellulase, while only D. rugosa PCD3-6 exhibited protease and phytase activities. Therefore, D. rugosa (strains PCD3-6, PCD93-19, PYJ93-7, PYJ93-16, PCJ94-4, PCJ94-6, PCJ94-10, PYD100-8, CTD93-1, CTD93-2, CTJ93-5, CTJ93-6, and YTJ97-4) and P. kudriavzevii (strains PYI3-3, PCI3-4, PCJ90-7, and PYI91-15) were proposed as potential probiotic yeasts. They may be beneficially applied as feed additives to improve nutritional value and quality, thereby enhancing animal performance and health. Moreover, this work indicated useful preliminary results that could be a baseline for future research in human probiotic applications.

In situ Single-Pass Perfused Rabbit Intestinal Study sample Analysis of Ractopamine by UPLC

In recent years, the prediction of oral drug absorption in humans has been a challenge for researchers and many techniques for permeability studies have been developed for several purposes, including biowaiver processes. The Single-Pass Intestinal Perfusion (SPIP) method performed in rats can provide permeability results closest to in vivo condition. he use of SPIP in rats and calculations for absorption prediction in humans may indicate the transport mechanisms and/or pre-systemic metabolism involved on permeation processes of drugs, since this model is the closest to in vivo conditions. Ractopamine hydrochloride is a commercial beta-adrenergic agonist commonly used as a dietary supplement in cattle production for improved feed efficiency and growth promotion. Currently, regulatory target tissues (as approved in the New Animal Drug Application with Food and Drug Administration) for ractopamine residue testing are muscle and liver. The aim of the study is to develop and validate an UPLC assay method to determine whether detectable and quantifiable levels of perfused ractopamine in small intestine of cattle or not. The separation is carried on Acquity UPLC BEH C18 (150mm x 2.1mm, 1.7 µm) reversed phase column using acetonitrile and 0.01M ammonium formate as the mobile phase in gradient mode at flow rate of 0.5mL/min. The precision, accuracy, extraction recovery, matrix effect, and stability meet the requirements of the guiding principles. In order to verify the sensitivity and selectivity of the method in a real-time situation, the developed method is successfully applied to a perfusion study in small intestine of rabbit and the samples were analyzed by Empower software.

Isolation, Identification and Detection of Campylobacter Fetus from Dairy & Poultry Products and Development of PCR

Campylobacters spp. are mainly responsible for gastrointestinal infections. The main source of infection is the consumption of contaminated or undercooked meat (especially poultry products), unpasteurized dairy products and untreated water. It is a type of zoonotic disease where it spreads through feces and mouth. Campylobacter species are widespread among warm-blooded animals, so they are found in food animals such as poultry, cattle, pigs, sheep and ostriches. Therefore, animal samples were used in this study. The most common causes of this infection are Campylobacter jejuni (C.jejuni), Campylobacter coli (C.coli) and Campylobacter fetus (C. fetus). Molecular techniques such as multiplex polymerase chain reaction (mPCR) have been optimized to detect these species. Campylobacter fetus was initially divided into two subspecies to reconcile significant epidemiological, clinical, and biochemical differences between the two similar taxa, which were later shown to be highly related at the genomic level. Campylobacter fetus occurs mainly in the intestines of cattle and sheep and can cause spontaneous abortion in both animals.

Adult Bovine-Derived Small and Large Intestinal Organoids: In Vitro Development and Maintenance.

Recent progress in bovine intestinal organoid research has expanded opportunities for creating improved in vitro models to study intestinal physiology and pathology. However, the establishment of a culture condition capable of generating organoids from all segments of the cattle intestine has remained elusive. Although previous research has described the development of bovine jejunal, ileal, and colonic organoids, this study marks the first report of successful bovine duodenal and rectal organoid development. Maintenance of these organoids through serial passages and cryopreservation was achieved, with higher success rates observed in large intestinal organoids compared to their small intestinal counterparts. A novel approach involving the use of biopsy forceps during initial tissue sampling streamlined the subsequent tissue processing, simplifying the procedure compared to previously established protocols in cattle. Additionally, our study introduced a more cost-effective culture medium based on Advanced DMEM/F12, diverging from frequently used commercially available organoid culture media. This enhancement improves accessibility to organoid technology by reducing culture costs. Crucially, the derived organoids from jejunum, ileum, colon and rectum faithfully preserved the structural, cellular, and genetic characteristics of in vivo intestinal tissue. This research underscores the significant potential of adult bovine intestinal organoids as a physiologically and morphologically relevant in vitro model. Such organoids provide a renewable and sustainable resource for a broad spectrum of studies, encompassing investigations into normal intestinal physiology in cattle and the intricate host-pathogen interactions of clinically and economically significant enteric pathogens.

Optimisation of adult Anopheles funestus blood-feeding on an artificial membrane feeding system

Malaria is one of the most severe vector-borne diseases caused by Plasmodium parasites and transmitted by Anopheles mosquitoes. Laboratory-reared anophelines are essential to advance research needed to reduce or eliminate malaria. The success of laboratory rearing as well as studies on parasite-mosquito transmission, is advanced by using an artificial membrane feeding systems. These require the optimisation of mosquito feeding to ensure that an optimal number of mosquitoes feed, thereby enabling successful reproduction or research sample sizes. In this study, various parameters such as the type of artificial membrane, density of adults in the feeding cup, age of the mosquito, duration of starvation, method of starvation, the volume of blood meal, duration of feeding, feeding in the light or dark and the effect of lactic acid were evaluated to determine their impact on the feeding rate of a main African malaria vector, Anopheles funestus. By optimising the artificial membrane feeding parameters, an increase in the feeding rate of the An. funestus mosquitoes was observed. The results obtained from these parameters increased the feeding rate of An. funestus above 50%. However, feeding rates were not significantly increased by the type of membrane, mosquito density, the volume of blood meal, duration of feeding and the addition of lactic acid to the cattle intestine membrane. Therefore, this study provides information on suitable conditions for adult mosquito feeding that allows for successful laboratory rearing and colony maintenance. Furthermore, it provides additional information for research studies that are dependent on blood-feeding, such as transmission blocking studies, endectocide studies etc.

Distribution of Toxocara vitulorum in Cattle of Ağrı Region

Parasitizing in the small intestines of cattle, buffalo and bison, Toxocara vitulorum is seen in countries with humid tropical and subtropical climates, including Türkiye. Adult parasites can cause digestive disorders such as loss of appetite, weakness, diarrhea or constipation, and sometimes death, especially in calves. This study was planned to investigate the distribution of T. vitulorum in cattle of the Ağrı region. In this study, stool samples of the size of a walnut were taken from the rectum of 200 calves 0-6 months old and 200 cattle older than 1 year in the province of Ağrı and its districts, and the preparations were prepared using the saturated zinc sulfate flotation method and the preparations were examined under a light microscope. T. vitulorum eggs were detected in 70 (35%) of 200 calves aged 0-6 months and 21 (10.5%) of 200 cattle over one year old. The difference in positivity between cattle over one year old and calves was statistically significant (p=0.001). With fecal examination ın 22 of the calves (11%), Trichostrongylidae spp. eggs were detected. It was determined that the prevalence of T. vitulorum in calves was high in Diyadin, Eleşkirt, Doğubayazıt, Hamur, Taşlıçay, Tutak and Patnos districts of Ağrı province. We believe that larger-scale studies should be conducted on the spread of this parasite, which causes significant low yields in cattle breeding.

Growth kinetics and fitness of fluoroquinolone resistant and susceptible Campylobacter jejuni strains of cattle origin.

Human enterocolitis is frequently caused by the Gram-negative microaerobic bacterium Campylobacter jejuni. Macrolides (e.g., erythromycin) and fluoroquinolones (FQs) (e.g., ciprofloxacin) are the preferred antibiotics for the treatment of human campylobacteriosis. Rapid emergence of FQ-resistant (FQ-R) Campylobacter during treatment with FQ antimicrobials is well known to occur in poultry. Cattle is also an important reservoir of Campylobacter for humans, and FQ-R Campylobacter from cattle has become highly prevalent in recent years. Even though the selection pressure may have contributed to the expansion of FQ-R Campylobacter, the actual impact of this factor appears to be rather low. In this study, we examined the hypothesis that the fitness of FQ-R Campylobacter may have also played a role in the rise seen in FQ-R Campylobacter isolates by employing a series of in vitro experiments in MH broth and bovine fecal extract. First, it was shown that FQ-R and FQ-susceptible (FQ-S) C. jejuni strains of cattle origin had comparable growth rates when individually cultured in both MH broth and the fecal extract with no antibiotic present. Interestingly, FQ-R strains had small but statistically significant increases over FQ-S strains in growth in competition experiments performed in mixed cultures with no antibiotic present. Lastly, it was observed that FQ-S C. jejuni strains developed resistance to ciprofloxacin more readily at high initial bacterial cell density (107 CFU/mL) and when exposed to low levels of the antibiotic (2-4 μg/mL) compared with that at a low level of initial bacterial cell density (105 CFU/mL) and exposure to a high level of ciprofloxacin (20 μg/mL) in both MH broth and the fecal extract. Altogether, these findings indicate that even though FQ-R C. jejuni of cattle origin may have a slightly higher fitness advantage over the FQ-S population, the emergence of FQ-R mutants from susceptible strains is primarily dictated by the bacterial cell density and the antibiotic concentration exposed under in vitro condition. These observation may also provide plausible explanations for the high prevalence of FQ-R C. jejuni in cattle production due to its overall fit nature in the absence of antibiotic selection pressure and for the paucity of development of FQ-R C. jejuni in the cattle intestine in response to FQ-treatment, as observed in our recent studies.

A systematic review of tapeworms (Plathelminthes, Cestoda) of Columba livia

After conducting research and analyzing literature data, it was discovered that Iraq is home to tapeworms belonging to four distinct families, namely Davaineidae, Hymenolepididae, Dilepididae, and Anoplocephalidae. Within these families, several species of tapeworm have been identified in Iraq, including Cotugnia intermedia, Raillietina tetragona, Raillietina cesticillus, Raillietina echinobothrida, Aporina delafondi, and Davainea proglottina. Cotugnia intermedia is a tapeworm commonly found in the intestines of small rodents and is transmitted to cats and dogs through the ingestion of infected rodents. Raillietina tetragona and Raillietina cesticillus are two species of tapeworm that commonly infect chickens, while Raillietina echinobothrida is found in pigeons and doves. Aporina delafondi is a tapeworm that has been found in the intestines of cattle, sheep, and goats in Iraq. The adult worms attach to the host’s intestinal wall and can cause intestinal inflammation and blockages. Finally, Davainea proglottina is a tapeworm that infects the intestines of domestic and wild carnivores, including dogs, cats, and foxes. Infection occurs through ingestion of infected intermediate hosts such as rodents, birds, and reptiles. Identifying these tapeworm species in Iraq is important for developing effective control and prevention strategies, as many of these parasites can cause significant health problems in animals and humans. Proper sanitation and hygiene practices, as well as deworming programs for domestic animals, are important measures that can help to reduce the spread of these tapeworms and their associated diseases.

Prevalence and molecular study of moniezia species isolate from cattle in Wasit Province, Iraq

Moniezia spp. parasitize the intestines of ruminants causing Monieziasis, M. expansa and M. benedeni (Cyclophyllidea: Anoplocephalidae) are large species of tapeworm that occurs in sheep and cattle and inhabits the small intestine.The current study was conducted during the period from December 2020 to August 2021 for detect the infection of Moniezia parasite in cattle using the traditional and molecular diagnostic methods as well as for confirmation of Moniezia spp. in cattle by the phylogenetic analysis. The samples fresh adult worms Moniezia were collected from intestines of cattle the locations of samples were obtained from abattoirs in Wasit province and also regarding months, sexes and ages affecting on infection prevalence rate.Results of microscopically, the samples fresh adult worms Moniezia spp. were collected from intestines of 125 cattle and stained with semichon's acid carmine and examined by light microscope, the results were revealed on 6 (4.8%) positive. Concerning to Moniezia spp. have been identified and classified out of 6 from cattle showed that M. expansa and M. benedeni 3 (50%) and 3(50%)respectively, without a significant difference.

Cotylophoron panamensis (Trematoda: Paramphistomidae) in Mazama americana (Artiodactyla: Cervidae) free-living in remote areas in the Peruvian Amazon.

Trematodes of the genus Cotylophoron cause mucosal lesions in pre-stomachs and in small intestine in cattle, generating losses in animal production. In the Northeastern Peruvian Amazon, the presence of Cotylophoron panamensis in the 16.7% (4/24) in free-living red brocket deers (Mazama americana) in areas distant from livestock farming suggests the red brocket is a natural wild host of C. panamensis.

Wickerhamiella martinezcruziae f. a., sp. nov., a yeast species isolated from tropical habitats.

Four yeast isolates with an affinity to the genus Wickerhamiella were obtained from beach sand, a marine zoanthid and a tree exudate at different localities in Brazil. Two other isolates with almost identical ITS and D1/D2 sequences of the large subunit rRNA gene were isolated from the small intestine of cattle and a grease trap in Thailand. These isolates represent a novel species phylogenetically related to Wickerhamiella verensis, Wickerhamiella osmotolerans, Wickerhamiella tropicalis, Wickerhamiella sorbophila and Wickerhamiella infanticola. The novel species differs by 15-30 nucleotide differences from these species in the D1/D2 sequences. The name Wickerhamiella martinezcruziae f.a., sp. nov. is proposed. The holotype of Wickerhamiella martinezcruziae sp. nov. is CBS 16104T. The MycoBank number is MB 839328.

Effects of Ionophores on Ruminal Function of Beef Cattle.

Simple SummaryIonophores are an important nutritional tool used to manipulate ruminal fermentation dynamics and improve the efficiency and performance of beef and dairy animals. Ionophores are carboxylic polyether antibiotics naturally produced by an occurring strain of Streptomyces spp. Ionophores modulate the ruminal environment by targeting and altering the metabolism of Gram-positive bacteria, resulting in an increased concentration of ruminal propionate and a reduced acetate concentration. Another pronounced effect of ionophores is the mitigation of ruminal proteolysis and the consequent reduction in ammonia synthesis. The purpose of this review is to highlight the impacts of ionophores on ruminal fermentation, leading to an improvement in the efficiency and performance of beef cattle.Ionophores have been widely used in the beef and dairy industry for decades to improve feed efficiency and performance by altering ruminal fermentation dynamics, increasing the level of propionate. Ionophores can also reduce ruminal proteolysis and ammonia synthesis, thus increasing the influx of protein into the small intestine in cattle, leading to improvements in performance and efficiency responses. Ionophores indirectly impact ruminal methanogenesis by decreasing the substrate used to produce methane. Despite the consistent benefits of using ionophores in cattle nutrition, their utilization is under public scrutiny due to concerns related to microbial adaptation. However, there is inconsistent evidence supporting these concerns, whereas ionophores are still an important dietary tool to enhance productivity and profitability in beef production systems.

Avaliação da proliferação celular de clostridium perfringens em intestino delgado de ruminantes / Evaluation of cellular proliferation of clostridium perfringens in small intestine of ruminants

This study evaluated the isolation, molecular typing and celular proliferation of C. perfringens in small intestine of cattle and sheep distributed in case and control groups. In the case group, out of the 61 bovine samples analyzed, C. perfringens was isolated in 29 (47.54%) and in 4 (33.34%) from 12 sheep samples. In the control group, we did not isolate the microorganism in the bovine samples (73 samples), however it was possible in 5 (20.83%) out of the 24 sheep samples. All isolates (100%) were classified as type A, and cell quantification showed that in control group, all cattle samples resulted in 10 CFU/g of intestinal content while in the study group presented a median value of 104 CFU/g, varying from 10 CFU/g through 108 CFU/g. In ovine group there was a clear divison of values between the two groups. Molecular typing corroborates with other studies regardin the epidemiological importance of type A in enterotoxaemia in ruminants, thus cellular quantification data allow us to conclude that healthy animals present a basal level of C. perfringens lower than 10 CFU/g that does not allow its isolation.

Changes in the structural components of the mucous membrane and plasma proteins of small intestine enterocytes in fetuses of cattle

The interrelationships of morphological changes of the mucosa of the large intestine with the dynamics of enzyme activity at different poles of enterocytes of cattle during the fetal period of ontogenesis are shown and analyzed. The work was performed on Holstein cattle aged two to nine months. The activity of hydrolytic and transport enzymatic systems in the basolateral membrane of cattle enterocytes depends less on the morphometric parameters of the large intestine than in the apical membrane. It is proved that during the fruitful period of ontogenesis there are dynamic transformations of enzyme systems of enterocytes of the large intestine of cattle which are connected with morphological changes of the mucous membrane of the large intestine. The activity of hydrolytic and transport enzymes in different domains of the plasma enterocytes of fetal cattle is interrelated with the morphometric parameters of the colon, in particular. the activity of alkaline phosphatase, γ-glutamyltransferase, Na+, K+-ATPase, Ca2 +, Mg2 +-ATPase and Mg-ATPase in the apical membrane of enterocytes is inversely related to the thickness of the intestinal wall with villi and the mucous membrane of the cavity (P ≤ 0.01–0.001). A significant correlation was found between the activity of alkaline phosphatase, γ-glutamyltransferase, Na+, K+-ATPase, Ca2 +, Mg2 +-ATPase and Mg-ATPase in the apical membrane of enterocytes with villi height (P ≤ 0.05). The activity of hydrolytic and transport enzymatic systems in the basolateral membrane of bovine fetal enterocytes depends to a lesser extent on the morphometric parameters of the large intestine than in the apical membrane. In particular, only the activity of alkaline phosphatase and γ-glutamyltransferase is inversely related to the thickness of the intestinal wall with villi and the mucous membrane of the jejunum with villi (P ≤ 0.01–0.001). The activity of transport enzymes is directly related to the morphometric parameters of the colon (P ≤ 0.01). The activity of these enzymes in the basolateral membrane does not significantly depend on the width of the villi, but is dependent on their height.

Industrial Biotechnology is Firing on All Cylinders

Industrial Biotechnology is Firing on All Cylinders

Cross Reaction of Haemonchus contortus Protein with Toxocara vitulorum Anti-L2 Serum Using Western Blot Technique

In the adult stage, Haemonchus contortus worms infect the abomasum host causing anemia and even death in animals. However, identifying the H. contortus protein can be used as a reference for the diagnosis of diseases. The diagnosis is performed by serological cross-reaction between H. contortus protein and anti-L2 Toxocara vitulorum (T. vitulorum) serum using the western blot technique. The main purpose of the current research was to identify the cross-reaction between H. contortus proteins and anti-L2 T. vitulorum serum using the western blot technique. T. vitulorum worms were collected from the intestine of cattle and H. contortus worms were collected from the abomasum of goats. The first step was making antibodies by oral infection of rats with infective eggs (L2) of T. vitulorum. The blood was taken 21 days after infection. Then, the blood was centrifuged at 1500 rpm for 10 minutes to get the serum. The second step was making homogenates from the whole worm extract of H. contortus. After crushing the worms, it was centrifuged at 5000 rpm for 15 minutes and the supernatant was taken. The supernatant was then analyzed using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) with coomassie brilliant blue staining. The third step was the analysis of H. contortus protein with serum anti-L2 T. vitulorum using the western blot technique. From the H. contortus homogenates analysis using SDS-PAGE, 16 protein bands were obtained. The cross-reactions were 141.3, 81.3, 64. 6, 51.3, 46.8, and 38 kDa. The data from cross-reactions suggested that the H. contortus protein cannot be used as a diagnostic material. It is serologically Haemonchosis because it caused false positives with diagnostic Toxocariasis.

Modelling transmission and control of Mycobacterium avium subspecies paratuberculosis within Irish dairy herds with compact spring calving

Paratuberculosis is a chronic bacterial infection of the intestine in cattle caused by Mycobacterium avium subspecies paratuberculosis (Map). To better understand Map transmission in Irish dairy herds, we adapted the French stochastic individual-based epidemiological simulation model to account for seasonal herd demographics. We investigated the probability of Map persistence over time, the within-herd prevalence over time, and the relative importance of transmission pathways, and assessed the relative effectiveness of test-and-cull control strategies. We investigated the impact on model outputs of calf separation from cows (calves grazed on pasture adjacent to cows vs. were completely separated from cows) and test-and-cull. Test-and-cull scenarios consisted of highly test-positive cows culled within 13 or 4 weeks after detection, and calf born to highly test-positive cows kept vs removed. We simulated a typical Irish dairy herd with on average 82 lactating cows, 112 animals in total. Each scenario was iterated 1000 times to adjust variation caused by stochasticity. Map was introduced in the fully naive herd through the purchase of a moderately infectious primiparous cow. Infection was considered to persist when at least one infected animal remained in the herd or when Map was present in the environment.The probability of Map persistence 15 years after introduction ranged between 32.2–42.7 % when calves and cows had contact on pasture, and between 18.9–29.4 % when calves and cows were separated on pasture. The most effective control strategy was to cull highly test-positive cows within four weeks of detection (absolute 10 % lower persistence compared to scenarios without control). Removing the offspring of highly test-positive dams did not affect either Map persistence or within-herd prevalence of Map.Mean prevalence 15 years after Map introduction was highest (63.5 %) when calves and cows had contact on pasture. Mean prevalence was 15 % lower (absolute decrease) when cows were culled within 13 weeks of a high test-positive result, and 28 % lower when culled within 4 weeks.Around calving, the infection rate was high, with calves being infected in utero or via the general indoor environment (most important transmission routes). For the remainder of the year, the incidence rate was relatively low with most calves being infected on pasture when in contact with cows. Testing and culling was an effective control strategy when it was used prior to the calving period to minimize the number of highly infectious cows present when calves were born.

Moniezia, Sp is found inside Cow Intestines Slowing down the Growth of the Cattle

Cows are the most recognized ruminants in the tropics. Traditionally, ruminant animal feed comes from agricultural waste which is high coarse fiber feed. About 60% to 75% of cattle feed in the form of crude fiber consists of carbohydrates such as cellulose, hemicellulose, pectin, and lignin. The pattern of cattle husbandry management in Indonesia has led to a more modern system. Currently, cattle grazing is not only in grasslands but also in landfills (TPA). Cows that are grazed in TPA get their food intake from fermented organic waste, but there are side effects that are not good for the health of cows, especially parasites that live in damp places. This study aimed to identify Moniezia, Sp in the intestines of cattle in Indonesia. This research method is by using PCR, sequencing, and alignment of amino acids using Blast, The amino acid Monieziabenedeni in bovine intestines was analyzed using Swiss Prot software to determine the three-dimensional structure. Prediction of B cell epitope using IEDB software, allergen levels, and toxicity using AllerTop and ToxinPred software. From the results of the research conducted, it was found that Monieziabenedini was found in the intestines of cows. More is needed regarding the characteristics of Monieziabenedini in bovine intestines, especially those related to pathogenesis and potential as a vaccine.

PSVI-14 Differentially expressed microRNAs with potential regulatory roles in ileum of Holstein cows with subclinical Johne’s disease

Abstract Paratuberculosis or Johne’s disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic inflammatory disorder of the intestinal tract of ruminants. The mechanisms of JD pathogenesis remain unclear but MAP can cause chronic inflammatory lesions in the terminal small intestine of cattle. Using microRNA (miRNA) transcriptome analysis, this study investigated the impact of MAP infection on miRNA expression and their potential regulatory roles in JD progression. Ileal tissues were collected from MAP infected Holstein cows (n = 5) and healthy cows (n = 5). Purified total RNA were used to construct libraries followed by next generation sequencing, bioinformatic processing using a standard pipeline, differential miRNA expression analysis with DESeq2 software, and functional enrichment with ClueGo app in Cytoscape. A total of 269 miRNAs were expressed in ileal tissue including three abundantly expressed miRNAs (Bta-miR-143; 54.05% of total read counts), bta-miR-145 (11.54%) and bta-miR-192 (4.31%)). Twenty miRNAs were significantly (P < 0.05) differently expressed (DE) when comparing ileal tissues from MAP infected and uninfected cows. Bta-miR-383 (fold change = -12.93) and bta-miR-1842 (fold-change = 8.17) were the most down- and up-regulated miRNAs, respectively. Amongst the DE miRNAs, bta-miR-146a, bta-miR-146b and bta-miR-393 are inflammation-related miRNAs and considered potential biomarkers of bovine mastitis and other diseases, while bta-miR-370 and bta-miR-383 are associated with lipid metabolism. The predicted target genes of DE miRNAs were enriched (P < 0.05) for gene ontology terms related to regulation of metabolic, cell and developmental processes, and immune system development. KEGG pathway analysis revealed potential involvement in immune regulation (e.g. T-cell receptor signaling and C-type lectin receptor signaling pathways) and pathways in cancer were also enriched (P < 0.05). Our results indicate that some DE miRNAs play important roles by regulating metabolism and immune processes during JD and provide DE miRNAs that might form targets for future development of intervention measures.

LYSIS TEST AND BIOASSAY FOR EXPERIMENTAL INFECTION OF CALVES WITH BUXTONELLA TO DETERMINE THE EFFECTIVENESS OF THE DRUG CYSTODESIS

To combat the parasitic protozoa of cattle and, above all, eimeriosis, preparations have been proposed that are effective against both exogenous and endogenous stages. Such studies have not been conducted before in relation to cattle buxtonellosis. The lysis test was performed with various concentrations of the cystodesis drug, and the bioassay was carried out to determine the effectiveness of the drug for disinvasion. The efficacy of disinvasion in the appointment of different concentrations of the drug cystodesis, as well as 4% concentration of the basic drug phenol was determined based on the percentage reduction in the allocation of cysts Buxtonella after exposure to the above-mentioned drugs and concentrations compared to calves infected controls, which were prescribed 200 cysts / ml. Intensity of the drug Cystodesis in 3% concentration was 97.9%, and in concentrations of 4 and 5%, the drug showed 100% efficiency. Conducted production tests showed that the complex preparation of cystodesis 4% for disinvasion of environmental objects in cattle farms in a dose of 0.5 l per 1 m2 at an exposure of 2 hours showed 92.32% intensity against cysts Buxtonella. The proposed tool, which includes the use of a complex concentrated drug cystodesis against cysts Buxtonella, provides high intensity efficacy for cattle buxtonellosis. Studies of domestic and foreign scientists have established that every cattle-breeding farm that practices stall-walking maintenance with the use of pastures is unfavorable for parasitic diseases and, above all, for parasitic protozoa, among which buxtonellosis is most common due to the extensiveness of invasion. According to the literature in the intestines of cattle of different ages, one type of ciliates of Buxtonella sulcata parasitizes.To combat the parasitic protozoa of cattle and, above all, eimeriosis, preparations have been proposed that are effective against both exogenous and endogenous stages. Such studies have not been conducted before in relation to cattle buxtonellosis. Based on the above, the problem of cattle buxtonellosis sets the task of researchers to develop measures to combat both exogenous and endogenous stages of invasion development and, above all, to develop a method for disinvasion of environmental objects against cysts Buxtonella.