The primary focus of the current study was to determine the potential benefits of supplementing sheep diet with white button mushrooms (WBM) in terms of growth, health and the kinetics of systemic ...CD4+ CD8+ memory T lymphocytes in lambs. Forty-five female lambs (Lika breed) were divided into three groups: A – the control group fed on a free-range pasture for the 222 days of the experiment, while groups B and C were housed in a separate facility for 42 days and fed either a commercial feed mixture (FM) or a FM supplemented with 15% of freshly prepared WBM, respectively, and ad libitum forage. For the remaining 180 days of the experiment, both groups (B and C) of lambs were kept free-range and fed pasture only. The lambs were monitored daily starting on Day 0 (or 90 days of age) before the treatments, weighed and blood sampled on Days 0, 21, 42 and 222, and were clinically observed for the incidence/severity of diarrhea and/or other signs of disease. In addition to morbidity, mortality was also monitored, and dead lambs were examined for gross pathology changes. The lambs fed FM supplemented with WBM (group C) had significantly higher body weight gain (P<0.05) on Days 42 and 222. They were neither diarrheic nor had any mortality cases throughout the experiment. Also, these lambs had a significantly increased (P<0.05) proportion of CD4+CD8+ T cells on Days 42 and 222. The data obtained supported our assumption of the efficacy of dietary WBM in the immunostimulation of CD4+CD8+ memory T lymphocytes in lambs, resulting in protection against on-farm diarrhea and providing an increased growth rate.
Pregnancy termination is a required procedure in companion animal practice. In healthy bitches with confirmed pregnancy, good results are obtained with the combination of prostaglandin F
2
and ...dopamine agonist, followed by regular sonographic examination until confirmed abortion. The aim of this study was to establish a simple and easily applicable procedure, with different dynamics of application of dinoprost and cabergoline. Dinoprost was administered intramucousally in the vestibule of vagina. Twenty bitches were divided into four equal groups. Group A received dinoprost and cabergoline daily; group B received dinoprost every 48 h and cabergoline daily; group C received dinoprost daily and cabergoline every 48 h, and group D received both cabergoline and dinoprost every 48 h. The treatment lasted until abortion was sonographically confirmed. The pregnancy was successfully terminated in all bitches, and side effects appeared in 90% animals, though they were of strong intensity in only 15% of bitches. Dinoprost administered intramucousally was effective with fewer undesirable, strong, systemic side effects. Drug administration every 48 h also induced abortion, with negligible side effects, but with slightly longer duration of treatment. This combination could be interesting in cases with time-limited owners and in countries where other drugs are unavailable.
This study aimed to identify seasonal and age related variability in triglycerides (TRI), paraoxonase (PON), stimulated PON (PON I) and cholesterol (CHOL, HDL, LDL) in extensively reared Lička ...pramenka sheep. The study was conducted 30 randomly chosen non-gravid Lička pramenka sheep on an extensive farm, grouped into three age groups in the Karlovac County, Croatia. Blood was collected by venepuncture of the jugular vein, twice in one year - the first sampling in spring (April) and the second in autumn (October). The results showed that age had no significant effect on any of the investigated indicators, while seasonality significantly affected five out of the six parameters examined. Significantly higher levels of serum lipid components in autumn can be explained by the influence of the traditional way of rearing and feeding Lička pramenka sheep. Furthermore, the results of PON I in all three age groups showed that the sheep are well adapted to the environment and to the rearing system. Future research should be carried out on higher number of farms, and should include more sheep in order to determine objective benchmarks for serum lipids and the enzymes connected to those lipids.
The objectives of this study were to investigate the effect of the dog’s age, semen quality, and the duration and season of semen transit on whelping rate and litter size after insemination with ...transported chilled extended semen. The sperm rich fraction was collected from 43 dogs of 18 breeds, which were presented at the Clinic for chilled semen transport, in the period from 2017 to 2021. Immediately after collection, the total sperm concentration and count, motility, membrane integrity (HOS test), the percentage of live spermatozoa and sperm morphology (eosin nigrosin staining) were evaluated. The sperm rich fraction was centrifuged and diluted with Tris – fructose - citrate extender with the addition of 20% (v/v) egg yolk, then chilled and prepared for shipping. A dose consisted of at least 200x106 live, motile, morphologically normal spermatozoa. The data on the dog’s age, chilled sperm transit time, the season of transit, and the whelping rate and litter size after insemination were recorded. The whelping rate was 55.8% with a mean (±SEM) litter size of 4.71±0.58 pups. The total number of spermatozoa was higher in artificial insemination (AI) that resulted in whelping compared to unsuccessful AI (P<0.05). No difference was observed for any of other sperm quality parameters tested, such as the dog’s age or season of transit regarding whelping rate or litter size. Transit time significantly affected the whelping rate (P<0.01), at (mean±SEM) 21.50±1.28 and 37.00±5.59 h in successful and unsuccessful AI, respectively. In conclusion, analysis of the factors related to the dogs identified total sperm count and transit time as factors that significantly affected whelping rates in bitches inseminated with transported chilled extended semen.
Enterotoxigenic Escherichia coli (ETEC) strains expressing F4 and F18 fimbriae are the most common causative agents of post-weaning diarrhoea (PWD) in pigs. The growing global restriction on the use ...of antibiotics in food animals has encouraged research into the development of nutritional and feeding strategies as well as vaccination against PWD. The aim of this study was to evaluate the efficacy of a live oral F4ac+ F18ac+ non-ETEC vaccine candidate (VAC) to stimulate gut and systemic cellular immunity in 4-week old pigs over 5 weeks following immunization. The onset and duration of protective immunity against on-farm occurring PWD, growth performance, diarrhoea scoring and mortality, as well as the phenotypic proportions of immune cells, were determined. Faecal and ileal samples were taken for determining the microbial composition or phenotyping of naïve/memory T cells. Also, the effect of prebiotic supplement mannan oligosaccharide (MOS) in the prevention of small intestinal colonization by ETEC, and its potential adjuvanticity in combination with the vaccine (VAC+MOS) were assessed. The pigs supplemented with MOS or that received VAC had significantly higher body weight (BW) (P<0.05) on Day 14, whereas the VAC+MOS treated pigs had significantly lower BW on Day 35. Treatment with VAC+MOS resulted in considerably reduced clinical PWD, in particular the incidence and severity of diarrhoea and mortality. The total bacterial load in the ileum was much lower in the pigs from all 3 principal groups (MOS, VAC, and VAC+MOS) than in the control (CON) group (19 x 107, 17 x 107 and 12 x 107 vs. 23 x 108 CFU/mL, respectively) on Day 35. The pigs from the principal groups had significantly higher proportions of tested immune cells (P<0.05) on Days 28 and 35. The localization and frequency of naive CD45RA+ and memory CD45RC+ T lymphocytes indicated their different distribution patterns within particular tissue structures, such as the villi, crypts, epithelium, lamina propria and areas (interfollicular follicular and Peyer’s patches) of ileal mucosa. This may indicate their different functions in intestinal immune responses to intraluminal microbes and their products, vaccinal immunogens and/or immunomodulators/adjuvants. To conclude, active mucosal immunity is needed to protect pigs against PWD. Hence, oral vaccination of pigs against both F4 and F18 ETEC, in combination with prebiotic supplementation represents a sustainable, practical and effective approach in PWD control.
No safe and effective vaccine exists against porcine enterotoxigenic Escherichia coli (ETEC) strains, which are the etiological agents of post-weaning diarrhoea (PWD), economically one of the most ...significant diseases of swine, which encountered for major productive losses in swine industry worldwide. The current study was designed to evaluate: (1) efficacy of an oral bivalent F4ac+/F18ac+ non-ETEC live vaccine candidate (VACCINE) in stimulating systemic and intestinal cellular immunity in 4-week-old weaned pigs, (2) the onset and duration of protective immunity of weaned pigs against naturally occurring PWD during the period of 6 weeks following weaning, and (3) the dietary supplement potential of zeolite clinoptilolite (CPL), an antimicrobial mineral and/or immunomodulator/ vaccine adjuvant (VACCINE + CPL). The pigs immunized either with the VACCINE or its combination with dietary CPL had significantly increased body weight gain from Day 7 to Day 42 (P<0.05) of the experiment, as compared to the control pigs (CONTROLS). Conversely, the pigs that were only supplemented with CPL had mostly significantly lower (P<0.05) body weight. The pigs that received VACCINE + CPL were neither diarrheic nor were there any mortalities during the entire period of the experiment. On Day 42 the total bacterial load in the jejunum was much lower in the pigs from all three principal groups than in the CONTROLS (30 x 108 vs. 18 x 107 vs. 14 x 107 vs. 13 x 107 CFU/mL, respectively). Regarding CD4+ and CD8+ T cells, specific immunization with either VACCINE or with VACCINE + CPL stimulated a significantly higher proportion of these cells (P<0.05) from Day 7 to Day 42 of the experiment. Quite similar findings were obtained for CD21+ B cells, as their proportion was significantly elevated in the pigs treated with either VACCINE or VACCINE + CPL (P<0.05 and P<0.05, respectively). The pigs from the VACCINE + CPL group had a significantly higher proportion of CD45+ lymphoid cells than the pigs from the CONTROL group (P<0.05). Quite sparsely distributed CD45RA+ naïve T lymphocytes were observed in the jejunal lamina propria of the intestinal villi, within the Lieberkühn crypts (LC) and in the submucosa of the CONTROLS and CPL supplemented pigs. The treatment with the VACCINE induced moderate recruitment of CD45 RA+ cells within both the IFA and FA of the jejunal PP of the pigs 6 weeks following the vaccination. In the CONTROLS, CD45 RC+ memory T cells were not abundant and were mostly dispersed in the middle of the villous lamina propria, but only rarely adjacent to the basal membrane of the intestinal enterocytes. These cells were more frequent within the lamina propria of the villi in the CPL group of pigs than in the CONTROLS. Even more numerous CD45 RC+ memory T cells were observed in the villous lamina propria of the jejunum of the pigs from the VACCINE group. These cells were predominantly found in the villous lamina propria and in the IFA, but were less frequent in the FA of the jejunal PP of the pigs that received the VACCINE and CPL. Due to the nature of the disease, challenges in PWD vaccine development will continue to exist. Although our research approach was at least partially successful, developing a safe and immunogenically effective live oral vaccine against PWD that will provide solid protection and sustained cellular and humoral immune responses remains a significant challenge.
The aim of this study was to investigate the effect of dietary carob wholemeal (Ceratonia siliqua L.) through monitoring the changes in some blood parameters (RBC, LEU, CD45+, CD4+, CD8+, CD21+) on ...the growth and development of weaned pigs (Swedish Landrace and Yorkshire cross-breeds). Thirty pigs were divided into two groups. In the control group the pigs were fed with a standard feed mixture. In the second, the experimental group (EXP), the feed mixture was enriched daily with carob wholemeal in a dose of 40 g/kg diet. Both groups were fed ad libitum. At the end of the 42-day experiment the pigs in the EXP groups were on average 23% heavier. The total sugar content, total phenolic content, flavonoids, anthocyanins, tannins and total antioxidant content of the carob wholemeal was determined. Carob wholemeal contains high carbohydrate content (73%). The total phenolic content value was 9.98 + or - 0.18 mg GAE/g. The total flavonoid value was 6.56 + or - 0.19 mg GAE/g, or 65% of total polyphenols, from which it can be seen that antioxidant activity is correlated to the total phenolic content. The total anthocyanins content was 34.00 + or - 0.07 mg CE/g while the total tannin content was 658.75 + or - 18.75 microg TAE/g. The ABTS method values were 89.50 + or - 0.17 micromol TE/g sample while antioxidant activity determined by the DPPH method was 56.87 + or - 0.66 micromol TE/g sample. Carob supplementation did not affect the amount of red blood cells and leucocytes, but did affect the proportions of the proliferation rate of CD45+ lymphoid cells, CD4+ and CD8+ T cells and CD21+ B cells in peripheral blood. Between days 14 and 42 an increase in the proportions of CD45+, CD4+, CD8+, CD21+ cells (at pless than or equal to0.05 or pless than or equal to0.01, respectively) was observed. Dietary supplementation of weaned piglets with 4% carob wholemeal showed an advantageous or beneficial effect on the immunity and productivity of the weanlings. Key words: carob; diarrhoea; phytocompounds; immunology; pigs
Data concerning the morphometric parameters of sheep red blood cells (RBCs) obtained using computer-assisted image analysis have not yet been investigated, and there are no data on any analyses of ...ovine RBC subpopulations based on their morphometric parameters. The aims of this study are to determine the values of RBC haematological and morphometric size and shape parameters, to form groups according to the obtained values of haematological parameters; to determine the differences in RBC morphometric parameters between the formed groups, and to determine RBC subpopulations and their respective proportions in the formed groups. Thirty-six blood samples were collected from the jugular vein of clinically healthy Lika pramenka sheep, aged between 2 and 5 years. Haematological parameters including haemoglobin (HGB), haematocrit (HTC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and RBC distribution width were analysed using a haematology analyser. Haematological parameters were categorized into two groups: those with lower values or values below the physiological range (Groups 1) and groups with higher values or values above the physiological range (Groups 2). Morphometric parameters of RBCs were determined from stained blood smears using SFORM, a computer-assisted program. Significantly higher values of RBC area, outline, convex, minimal and maximal radius, as well as length and breadth were established in Groups 2 compared to Groups 1 of HGB, HCT, MCV, MCH, and MCHC, respectively. Based on the morphometric parameters of RBCs, three RBC subpopulations were obtained using principal component and cluster analysis: ES 1-the smallest and most elongated RBCs, ES 2-the biggest and most rounded RBCs, and ES 3-average size and shape RBCs. Significantly higher proportions of ES 2 and ES 3 subpopulations, as well as a significantly lower proportion of ES 1 subpopulation, were established in Groups 2 compared to Groups 1 of HGB, HTC, MCV, and MCH, respectively. It can be concluded that ovine RBC subpopulations, based on their morphometric parameters, can be obtained by using computer-assisted image analysis of RBC morphometry and multivariate statistical methods, including principal component and cluster analysis. RBC morphometry, including classification into subpopulations, could serve as a basis for future possibilities in the diagnostic interpretation of anaemic syndromes in veterinary medicine, especially in normocytic, macrocytic, and microcytic anaemias in sheep.
Sustav uzgoja i hranidbe janjadi, odnosno botanički sastav pašnjaka na kojima se janjad uzgaja ima značajan učinak na aromu i sastav janjećeg mesa („efekt teritorija“). Pri tomu znatan doprinos na ...formiranju arome janjetine imaju terpeni koji se izravno iz biljaka ugrađuju u životinjska tkiva preživača ili nastaju kao rezultat razgradnje klorofila pod utjecajem mikroflore buraga. Stoga je za istraživanja u okviru ovog rada, ali i samog HRZZ projekta “Inovativni funkcionalni proizvodi od janjećeg mesa“ (IP-2016-06-3685) od velikog interesa procjena brojnosti, gustoće i pokrovnosti aromatične svojte majčine dušice (Thymus pulegioides L.) s pašnjaka na području Velike Crkvine (Kordun). Rezultati rada pokazuju da na 20 ha površine procijenjenog pašnjaka majčina dušica (Thymus pulegioides L.) ima zamjetnu procjenu učestalosti od 19,2 % ± 4,54 % (95%-tne granice pouzdanosti 10,77 - 27,64 %) i mjerenu gustoću po kvadratnom metru od 0,26±0,09 (95%-tne granice pouzdanosti 0,09 - 0,44), odnosno procijenjenu gustoću od 2,5±0,53 (95%-tne granice pouzdanosti 1,52 - 3,48) stupnja prema Braun-Blanquetu skali s pet stupnjeva od ukupno determiniranih ljekovitih i aromatičnih vrsta krajem lipnja 2019. godine. Nadalje, njezina je pokrovnost procijenjena dronom iznosila 15,8±2,20 (95%-tne granice pouzdanosti 11,71 - 19,89), odnosno pripada 2a i 2b stupnju Braun-Blanquetove devetostupanjske skale.
Animal hairs are an apt surface for retention of forensic trace epithelial samples. The aim of this study was threefold: to evaluate different methods of sample collection (moistened and dry swabs) ...and DNA extraction (Chelex
100 method, Qiagen EZ1
DNA Investigator Kit), as well as to examine the morphological differences of hair fibres between two species (dog, sheep) and their ultimate impact on sample collection and processing. Our preliminary findings suggest that the use of EZ1
DNA Investigator Kit yields donor DNA profiles of higher quality. The results of different sample collection methods have shown intraspecific variations that require further investigation. The ability of retention and subsequent extraction of trace DNA appears to be similar between the two species, despite significant morphological differences between their coat hairs.
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