Učinak levamisola na brojnost crijevnih vrčastih stanica odbijene prasadi pokusno cijepljene protiv kolibaciloze

Postweaning colibacillosis (PWC) is an etiologically complex disease commonly induced by porcine F4ac+ enterotoxigenic Escherichia coli (ETEC) for which no effective vaccine is available. The objective of this study was to determine the nonspecific immunomodulatory effect of levamisole in combination with specific intragastric immunization of weaned pigs with a candidate F4ac+ non-ETEC oral vaccine on the population of intestinal goblet cells (GC). The pigs were immunized with F4ac+ non-ETEC strain, in combination with or without levamisole. Seven days after immunization the pigs were challenged with F4ac+ ETEC strain and 14 days following immunization they were euthanatized for sampling of specimens of the small intestine for immunohistochemistry and morphometric analyses. Samples of the ileum were tested for the presence of acidic and neutral carbohydrates, components of mucus produced and secreted by the intestinal goblet cells (GC). The volume density (VV) of the PAS+ and AB+/PAS+ GC was determined using the stereological point-counting method. The Vv of the ileal PAS+ GC was lowest (0.130 ± 0.075 mm3) in the pigs that were immunized with the vaccine candidate F4ac+ non-ETEC strain. Interestingly, AB+/PAS+ GC Vv were found to be highest (0.262 ± 0.091 mm3) in this group of pigs. The Vv of PAS+ GC was the largest (0.201 ± 0.064 mm3) in the negative control group of pigs. Pigs from the levamisole pretreated group were shown to have the lowest Vv (0.166 ± 0.051 mm3) of AB+/PAS+ GC. According to the data presented, there was no significant influence of nonspecific/specific immunization on the nonimmune defence mechanism of the intestinal mucosa, as measured by GC Vv in weaned pigs. Hence, it is likely that the production of mucus was not affected and that nonspecific protection of porcine intestines provided by mucus layer was not compromised following peroral immunization against PWC. Also, presumably the anti-inflammatory effect of levamisole was observed as the population of ileal mast cells was not considerably affected by non-specific/specific immunization and challenge infection.Kolibaciloza odbijene prasadi (KOP) etiološki je složena bolest, najčešće izazvana svinjskim F4ac+ enterotoksigenim sojevima bakterije Escherichia coli (ETEC), za koju nema djelotvorne vakcine. Namjera ovoga rada bila je da se utvrdi imunomodulacijski učinak levamisola, u kombinaciji sa specifičnom intragastričnom imunizacijom odbijene prasadi pokusnim vakcinalnim F4ac+ ne-ETEC sojem, na populaciju crijevnih vrčastih stanica (VS). Prasad je bila imunizirana F4ac+ ne-ETEC sojem u kombinaciji s levamisolom ili bez njega, sedam dana nakon toga bila je izazivački inficirana F4ac+ ETEC sojem, a 14 dana nakon imunizacije bila je usmrćena radi uzimanja uzoraka crijeva za imunohistokemijske i morfometrijske analize. Uzorci ileuma testirani su na prisutnost kiselih i neutralnih ugljikohidrata, sastavnica sluzi koju proizvode i izlučuju VS. Volumna gustoća (Vg) VS koje se boje kombinacijom alcijanskoga modrila (AM) i periodičnoga kiseloga Schiffovoga (PKS) reagensa određivana je pomoću stereologijske metode brojanja točaka. Vg ilealnih PKS+ VS bila je najniža (0,130 ± 0,075 mm3) u prasadi imunizirane pokusnim vakcinalnim F4ac+ ne-ETEC sojem. Međutim, zanimljivo je da je utvrđena najveća Vg AM+/PKS+ VS (0,262 ± 0,091 mm3) u prasadi iz te skupine. U prasadi iz negativne kontrolne skupine utvrđena je najveća Vg PKS+ VS (0,201 ± 0,064 mm3), dok je prasad iz skupine obrađene levamisolom i vakcinalnim ne-ETEC sojem imala najnižu Vv ukupnih (AM+/PKS+) VS (0,159 ± 0,023 mm3). Primjereno dobivenim rezultatima, nije utvrđen značajan utjecaj nespecifične/specifične imunizacije na neimunosne obrambene mehanizme crijevne sluznice vrednovane temeljem Vg i distribucije VS u odbijene prasadi. Stoga je vrlo vjerojatno da proizvodnja sluzi nije umanjena i da nespecifična zaštita crijeva prasadi slojem sluzi ne bi bila slabija nakon peroralne imunizacije protiv KOP. Također je zapažen potencijalni protuupalni učinak levamisola s obzirom na činjenicu da populacija ilealnih mastocita nije bila znatnije stimulirana nespecifičnom/specifičnom imunizacijom i izazivačkom infekcijom

Effect of levamisole on the number of intestinal goblet cells in weaned pigs experimentally vaccinated against colibacillosis

VALPOTIĆ: Effect of levamisole on the number of intestinal goblet cells in weaned pigs experimentally vaccinated against colibacillosis. Vet. arhiv 79, 543-553, 2009. Postweaning colibacillosis (PWC) is an etiologically complex disease commonly induced by porcine F4ac + enterotoxigenic Escherichia coli (ETEC) for which no effective vaccine is available. The objective of this study was to determine the nonspecific immunomodulatory effect of levamisole in combination with specific intragastric immunization of weaned pigs with a candidate F4ac + non-ETEC oral vaccine on the population of intestinal goblet cells (GC). The pigs were immunized with F4ac + non-ETEC strain, in combination with or without levamisole. Seven days after immunization the pigs were challenged with F4ac + ETEC strain and 14 days following immunization they were euthanatized for sampling of specimens of the small intestine for immunohistochemistry and morphometric analyses. Samples of the ileum were tested for the presence of acidic and neutral carbohydrates, components of mucus produced and secreted by the intestinal goblet cells (GC). The volume density (V) of the PAS V + and AB +/PAS+ GC was determined using the stereological point-counting method. The Vv of the ileal PAS + GC was lowest (0.130 ± 0.075 mm3) in the pigs that were immunized wit

Utjecaj hrane s dodatkom plemenite pečurke (Agaricus bisporus) na kinetiku promjena udjela CD4+ CD8+ T-limfocita u perifernoj krvi janjadi

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.Primarno usmjerenje ovog istraživanja bilo je utvrditi potencijalne pogodnosti dodavanja plemenite pečurke (PP) u hranu za ovce na rast, zdravlje i kinetiku sistemskih CD4+ CD8+ memorijskih T-limfocita u janjadi. Ukupno 45 janjadi ženskoga spola, pasmine lička pramenka, podijeljeno je u tri skupine. Kontrolna skupina A hranjena je na slobodnoj paši tijekom 222 dana pokusa, dok su skupine B i C bile smještene odvojeno u nastambi tijekom 42 dana i hranjene bilo komercijalnom krmnom smjesom (KS) bilo KS-om s dodatkom 15% svježe pripremljenog PP-a, uz dodatak voluminozne krme ad libitum. Preostalih 180 dana pokusa obje su skupine (B i C) janjadi držane u slobodnom uzgoju i hranjene samo ispašom. Janjad je svakodnevno nadzirana, počevši od nultog dana (ili 90. dana starosti) prije tretmana, vagana je te su uzimani i uzorci krvi 0., 21., 42. i 222. dan. Kliničkim pregledima ustanovljavana je pojavnost/jačina proljeva odnosno drugih znakova bolesti. Osim morbiditeta, praćen je i mortalitet, a uginula je janjad bila pregledana na patoanatomske i histopatološke promjene. Janjad hranjena KS-om s dodatkom PP-a (skupina C) imala je znakovito veći prirast tjelesne mase (P<0,05) 42. i 222. dan. Nije imala proljeva niti su zabilježeni slučajevi uginuća tijekom pokusa. Također, ta je janjad imala znakovito povećan (P<0,05) udio CD4+CD8+ T-stanica 42. i 222. dan pokusa. Dobiveni podaci potvrđuju našu pretpostavku o učinkovitosti PP-a kao dodatka hrani u imunostimulaciji CD4+CD8+ memorijskih T-limfocita u janjadi. Navedeno je rezultiralo zaštitom janjadi od proljeva i njihovim bržim rastom u farmskim uvjetima

Vrednovanje profilaktičkog potencijala imunizacije cjepivom neenterotoksigenog soja bakterije Escherichia coli (ne-ETEC) i pogodnosti kompetitivne ekskluzije dodatkom hrani manan-oligosaharida protiv infekcija izazvanih sojevima ETEC u odbijene prasadi

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.Enterotoksigeni sojevi bakterije Escherichia coli (ETEC), koji proizvode F4 i F18 fimbrije, najuobičajeniji su uzročnici dijareje nakon odbića (DNO) u prasadi. Rastuće globalno ograničavanje uporabe antibiotika u farmskih životinja usmjeruje istraživanja prema razvijanju nutritivnih i prehrambenih strategija, kao i prema cijepljenju protiv DNO-a. Cilj ovog istraživanja bio je vrednovanje učinkovitosti živog, oralnog F4ac+ F18ac+ ne-ETEC cjepiva-kandidata (VAK) u poticanju crijevne i sistemske stanične imunosti u prasadi u dobi od 4 tjedna, tijekom 5 tjedana nakon imunizacije. U pokusu su određivani početak i trajanje zaštitne imunosti od pojavnosti farmskog DNO-a, proizvodni rezultati, ocjenjivanje učestalosti i jačine dijareje te mortalitet i fenotipski udjeli imunosnih stanica. Uzimani su uzorci fecesa i ileuma za određivanje sastava mikrobiota ili za fenotipiziranje naivnih/memorijskih T-limfocita. Procijenjena je i učinkovitost prebiotika manan-oligosaharida (MOS), kao dodatka hrani, koji bi mogao uspostaviti kompetitivnu ekskluziju naseljavanja tankog crijeva sojevima ETEC, a mogao bi pokazati i adjuvantnost u testiranoj kombinaciji (VAK + MOS). Prasad hranjena dodatkom MOS-a u hrani, ili koja je primila VAK, imala je znakovito povećanu (P < 0,05) tjelesnu masu 14. dan pokusa, dok je prasad tretirana kombinacijom VAK-a i MOS-a imala znakovito nižu tjelesnu masu 35. dan pokusa. Tretman kombinacijom VAK-a i MOS-a rezultirao je znatno blažom kliničkom slikom DNO-a, napose u odnosu na pojavnost i jačinu dijareje te na mortalitet. Ukupno bakterijsko opterećenje u ileumu bilo je mnogo niže u prasadi iz sve tri pokusne skupine (MOS, VAK i VAK + MOS) od onog u kontrolnoj (KON) skupini (19 x 107, 17 x 107 i 12 x 107 prema 23 x 108 CFU/mL) 35. dan pokusa. Prasad iz pokusnih skupina imala je znakovito veće udjele testiranih imunosnih stanica (P < 0,05) 28. i 35. dan pokusa. Lokalizacija i učestalost naivnih CD45RA+ te memorijskih CD45RC+ T-limfocita pokazuju njihove različite obrasce smještanja u posebne tkivne strukture, kao što su crijevne resice, kripte, epitelij i lamina propria te područja u sluznici ileuma, što može specificirati njihove različite funkcije u crijevnim imunosnim odgovorima na intraluminalne mikrobe i njihove proizvode, vakcinalne imunogene i/ili imunomodulatore/adjuvanse. Zaključujemo da je radi zaštite prasadi od DNO-a nužno uspostaviti aktivnu mukoznu imunost. Stoga je oralno cijepljenje prasadi protiv F4 i F18 ETEC-a održiv, praktičan i učinkovit pristup u pronalaženju odgovarajućega bivalentnog, izrazito imunogenog i sigurnog cjepiva

BIOCLIMATE, KEEPING AND HOUSING OF ENGLISH FULL-BLOODED HORSES

Provedena su istraživanja stanja bioklime u jednoj od konjušnica zagrebačke regije s obzirom na klimatske uvjete ovog dijela Hrvatske, s ciljem da se dobiveni rezultati primjenjuju u izgradnji budućih konjušnica te popravljanju bioklimatskih uvjeta u već postojećim stajama za držanje i uzgoj konja. Tijekom jednogodišnjeg mjerenja (proljeće, ljeto, jesen i zima) praćeni su bioklimatski pokazatelji (temperatura zraka, relativna i apsolutna vlaga zraka i plinovi; ugljični dioksid - CO₂, amonijak -NH₃ i sumporvodik - H₂S). Nadalje, ukratko je prikazana tehnologija držanja, smještaja i iskorištavanja engleskog punokrvnjaka. Srednje temperature zraka nalaze se na samoj granici poželjnih temperatura (zimi 5°C, ljeti 28,5°C i u jeseni 22,0°C). Iz toga proizlazi da je termoizolacija promatranih nastambi za konje slaba. U promatranoj staji srednja relativna vlaga prelazi gornju poželjnu granicu. Razlog tome je loše rješenje provjetravanja i vlaženje hodnika prije metenja. Srednje vrijednosti za apsolutnu vlagu slijeće dobivene vrijednosti za relativnu vlagu. Srednje temperature zraka slijede temperaturu zraka,a oscilacije koje postoje mogu se pripisati slaboj termoizolaciji stropa i utjecaju brzine strujanja zraka. Srednje vrijednosti strujanja zraka, za sva mjerena godišnja razdoblja, kretala su se ispod poželjnih donjih granica, kao posljedica neadekvatne prirodne ventilacije konjušnice. Nazočnost plinova (CO₂, NH₃) u našim istraživanjima kretala se u granicama koje zastupaju i drugi autori, a H₂S nije registriran ni u jednom od naših mjerenja. Srednje najviše i najniže vrijednosti za navedene bioklimatske parametre prikazane su na slici 1, 2 i 3. Boksovi 3 x 3 m (9m²), na neki način odgovaraju toj kategoriji konja, ali su premali za ždrijebljenje. Pojilice i hranilice nisu na poželjnoj visini, što je posljedica dotrajalosti nabijenog poda od ilovače, pa je time konjima onemogučeno fiziološko uzimanje hrane. Tijekom istraživanja u konjušnici je bilo nazočno 13 grla engleskog punokrvnjaka, dok su ostala grla bili polukrvni jahačo konji.Bioclimate measurements were conducted in a stable in the Zagreb region, with regard to the climatic conditions of this pad of Croatia, with the purpose of applying the results in future stable construction and improvement oi conditions in existing horse stables. Nineteen riding horses were kept in the stable. Thirteen were English full-blooded horses, and the rest were halfbreeds. During one year period (spring, summer, autumn, winter) bioclimate indicators (air temperature, relative and absolute air humidity, average radiation temperature, relative and absolute air humidity, average radiation temperature, air speed, and gases CO₂, NH₃, and H₂S) were monitored and their average values presented for each season of the year. Furthermore, the technology of keeping, housing, feeding, and sport exploitation of English full-blooded horses was recorded

BIOCLIMATE, HEALTH STATUS AND WEIGHT GAIN IN FATTENING SWINE FED DIFFERENT DIETS

Istraživan je bioklimat u predtovu i tovu velike svinjogojske farme u području kontinentaine klime s alpskim utjecajem. Tijekom jedne godine po mjesecima praćeni su bioklimatski parametri: temperatura zraka, bzina strujanja zraka, relativna i apsolurna vlaga zraka, plinovi Co₂ i NH₃. Njihove najniže i najviše srednje vrijednosti prikazane su na slici 1, 2 i 3. U istraživanjima korišeni su trostruki križanci (švedski landras x veliki jorkšir x njemački landras). Opisana je ukratko tehnologija držanja i smještaja. Tijekom tova životinje su držane na prehrani kompletnim smjesama za predtov-tov. U prvom dijelu praćenja svinje,su hranjene tekućom hranom tijekom cijelog razdoblja, dok je u drugom dijelu praćenja, nakon rekonstrukcije objekata, u objekte uvedena oprema sistema "IBO". Nadalje, dat je prikaz proizvodnje tovljenika u razdoblju od 1975. do 1984. (slika 4) iz kojega se mogu vidjeti: prirast i konvenija hrane u kilogramima, prisilna klanja, ugibanja i gubitci u postotcima kao i početak i završetak rekonstukcije farme, te razdoblje provedene eradikacije dizenterije na farmi.Bioclimate studies were condusted in pre fattening and fattening units of a large swine farm, located in the continental climate zone influenced by alpine climate. Bioclimate parametres (air temperature, air speed, relative and absolute humidity, and gases CO₂ and NH₃) were monitored monthly during one year period. Minimal and average values were recorded. The management and housing technology of triple crossbreed (Swedish Landrace x Yorkshire x German Laandrace) is briefly described. During the fattening period the pigs were fed with complete pre fattening and fattening mixtures. In the first part of monitoring the pigs were fed with liquid feed. In the second part of monitoring, after the reconstruction of the facilities, they were fed the IBO system equipment. The results of production of fattening pigs in the period from 1975 to 1984 are also presented (growth rate and feed conversion, necessary slaughter, losses) as well as health status after the reconstruction of the farm and eradication of swine dysentery

Procjena kinetike CD45+ leukocita pune krvi tovnih pilića imuniziranih živim ili inaktiviranim cjepivom protiv newcastleske bolesti

Since insight into the effects of Newcastle disease vaccination on cellular immune responses has been insufficient to date, this study follows the leukocyte dynamics in the blood of fattening chickens inoculated against Newcastle disease with lyophilised live vaccine of the La Sota virus strain (experimental group E1) and/or inactivated vaccine produced from the same strain (experimental group E2). Blood was treated with labelled antibody for the CD45+ leukocyte marker and examined by flfl ow citometry. In bouth experimental a statistically signififi cant increase (P<0.01) of CD45+ cells was observed in relation to the control group of chickens, and the increase was more intensive and faster in the chickens vaccinated with lyophilised live vaccine than it was in chickens vaccinated with inactivated vaccine. The group treated with live vaccine had a signififi cantly increased number of CD45+ cells 6, 12, and 24 hr, and 7, 14, 21 and 28 days after inoculation. But on the 14th day after vaccination, a signififi cantly increased number of CD45+ cells (P<0.05) was found in the chickens inoculated with inactivated vaccine. It can be concluded that the lyophilised live vaccine was more efficient and cellular immunity was achieved sooner than with the inactivated vaccine during the observed period of 28 days after inoculation.Budući da su dosadašnje spoznaje o djelovanju cjepnog virusa newcastleske bolest na stanični imunosni odziv u peradi nedostatne, u ovom radu praćena je dinamika leukocita u punoj perifernoj krvi tovnih pilića cijepljenih protiv newcastleske bolest liofi liziranim živim cjepivom proizvedenim od soja La Sota virusa (pokusna skupina E1) i/ili inaktiviranim cjepivom proizvedenim od istog soja (pokusna skupina E2). U tovnih pilića cijepljenih liofi liziranim živim cjepivom ili inaktiviranim uočen je statistički značajan porast (P<0,01) CD45+ leukocita u punoj perifernoj krvi, pri čemu je taj porast bio intenzivniji i brži u pokusnoj skupini tovnih pilića koja je bila tretirana liofiliziranim živim cjepivom. Promatrajući obje pokusne skupine, skupina cijepljena živim cjepivom protiv newcastleske bolest 6., 12., 24. sata, 7., 14., 21. i 28 dana nakon cijepljenja je imala statistički značajno veću vrijednost za CD45+ leukocita. Međutim, 14. dana nakon cijepljenja znatan porast vrijednosti (P<0,05) za CD45+ leukocite ustanovljen je u skupini tovnih pilića cijepljenih inaktiviranim cjepivom. Može se zaključiti da je liofilizirano živo cjepivo pokazalo bolju učinkovitost i brže postizanje stanične imunosti od inaktiviranog cjepiva tijekom promatranog razdoblja od 28 tjedana nakon cijepljenja