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The idea of dual sire insemination in dog breeding is to give equal chances to both males to fertilize the eggs due to the specific oestrus cycle of the bitch. Two sexually mature, privately owned females Petit Basset Griffon Vendeen (bitch A and B) aged 3 to 5 years were subjected to dual- sire inseminations. Prior to insemination, the semen of stud dogs (n=5) was collected and evaluated for volume, concentration, total sperm count, motility, membrane integrity (HOS test), percentage of live spermatozoa and sperm morphology (eosin nigrosin staining). The time of insemination was based on serum progesterone (P4) concentrations, where P4 of >5–10 ng/mL was considered ovulation. Bitch A was inseminated in two oestrus cycles with fresh mixed semen from two males using the endoscopic transcervical insemination technique (TCI). Bitch B was inseminated in one oestrus cycle by laparoscopic intrauterine deposition of frozen thawed semen. Semen evaluation showed minimum deviations in all tested parameters between the chosen males on the days of insemination. Depending on sperm concentration and quality, the volume of ejaculates was adjusted, resulting in an equal number of motile spermatozoa from two males in each insemination, providing them an equal chance for fertilisation. Confirmation of pregnancy was carried out 24 days after insemination by ultrasonography. The whelping outcome was obtained directly from the owner of the bitches. Puppy blood was taken immediately after whelping from the umbilical vein into EDTA tubes. Blood samples were also obtained from the dam and both sires for DNA profiling. Parentage was determined for each dual-sired pup by using Thermo Scientific Canine Genotypes Panel 1.1. All inseminations resulted in pregnancy and whelping. A total of 14 puppies were born in three litters. Mixed parentage was determined in 1 of the 3 resultant litters (bitch A). In conclusion, dual sire insemination is a useful breeding tool providing the opportunity to obtain puppies from multiple genetic backgrounds in a single litter. However, as this method usually produces offspring from single father, optimal insemination protocol should be established for producing a litter of mixed paternity. more...

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, Svrha rada bila je istražiti utjecaj dobi psa, kvalitete sjemena te trajanja i sezone transporta ohlađenog sjemena na postotak štenjenja i veličinu legla nakon osjemenjivanja kuja ohlađenim, transportiranim sjemenom. U razdoblju od 2017. do 2021., od 43 psa 18 različitih pasmina koji su došli na kliniku s ciljem transporta ohlađenog sjemena, prikupljena je spermom bogata frakcija ejakulata. Neposredno nakon uzimanja ejakulata ocjenjena je koncentracija, ukupan broj spermija, pokretljivost, integritet membrane (HOS test), postotak živih spermija i morfologija spermija (bojenje eozin nigrosinom). Spermom bogata frakcija je centrifugirana i razrijeđena s Tris – fruktoza – citratnim razrjeđivačem s dodatkom 20% žumanjaka, nakon čega je uzorak ohlađen i pripremljen za transport. Doza za osjemenjivanje sadržavala je najmanje 200x106 živih, pokretnih, morfološki normalnih spermija. Zabilježeni su podatci o dobi pasa, tranzitnom vremenu, sezoni transporta te postotku štenjenja i veličini legla nakon osjemenjivanja transportiranim ohlađenim sjemenom. Od ukupnog broja osjemenjenih kuja, 55,8% se oštenilo s prosjekom (±SEM) od 4,71±0,58 šteneta po leglu. Ukupan broj spermija bio je viši kod osjemenjivanja koja su rezultirala štenjenjem nego kod neuspješnih osjemenjivanja (P more...

Globalno zatopljenje predstavlja direktnu i indirektnu prijetnju preživljavanju mnogih živih bića na zemlji. Uz direktne prijetnje koje donose prirodne katastrofe koje prate klimatske promjene poput uragana, tsunamija, poplava i suša, tu je i vrlo značajan utjecaj toplinskog stresa. Konji čija je sezona razmnožavanja direktno povezana s toplijim dijelom godine i sezonom najduljih dana, nalaze se pred fiziološkim izazovom. Njihova zona komfora je do 25°C a dobro podnose temperature do 30°C. Donedavno su temperature iznad 25°C tijekom 24 i više sati u umjerenom pojasu bile iznimna a danas su česta pojava. Stoga je vrlo vjerojatno da će razmnožavanje konja kao vrste u uvjetima globalnog zatopljenja predstavljati sve veći izazov kojem nije moguće doskočiti niti metodama asistirane reprodukcije jer je utjecaj toplinskog stresa na rasplođivanje konja kompleksan i sveobuhvatan. more...

Mastitis je među najčešćim bolestima na farmama mliječnih krava za čije se liječenje i prevenciju koristi najveća količina antibiotika. Učestala, neispravna ili nepotrebna uporaba antimikrobnih sredstava predstavlja veliku prijetnju ubrzanom razvoju antimikrobne rezistencije. Prijenos antimikrobno rezistentnih patogena na ljude moguć je putem mlijeka i mliječnih proizvoda te su propisane smjernice za racionalnu primjenu antimikrobnih sredstava u veterinarskoj medicini. U mliječnoj se industriji najviše antibiotika koristi prilikom zasušivanja krava u svrhu prevencije nastanka mastitisa. U isto vrijeme antibiotici se nepotrebno primjenjuje i u životinja koje nisu inficirane. Iz tog razloga sve više zemalja prelazi na model selektivnog zasušivanja krava. Antibioticima se zasušuju samo inficirane četvrti ili vimena krava, odnosno krave s povećanim rizikom za nastanak infekcije. Jedan od većih problema prilikom uvođenja selektivnog zasušivanja krava je odabir kriterija prema kojem će se odabrati krave koje će biti zasušene antibiotikom. Protokoli za odabir krava trebaju biti dovoljno precizni, jednostavni za interpretaciju, jeftini, sigurni, certificirani i dostupni za uporabu u svim dijelovima zemlje. Kriterij za odabir krava može se temeljiti na mikrobiološkoj pretrazi mlijeka, određivanju BSS prilikom zasušivanja, incidenciji kliničkih mastitisa u laktaciji, paritetu i njihovim međusobnim kombinacijama. Bakteriološka je pretraga mlijeka najpouzdaniji pokazatelj, ali se broj somatskih stanica na posljednjoj mužnji ističe kao pouzdan pokazatelj na temelju kojeg se može provesti selektivno zasušenje bez negativnih učinaka na proizvodnju. Granična vrijednost broja somatskih stanica kao učinkovit i održiv model za donošenje odluka iznosi 100.000 st/mL mlijeka za primiparne i 200.000 st/mL mlijeka za multiparne krave. Uporaba antibiotika za zasušenje samo kod inficiranih četvrti krava omogućava dodatno smanjenje primjene antibiotika. Iako je tijekom godina postignut značajan napredak u razumjevaniju epidemiologije, imunologije, dijagnostike i patogeneze intramamarnih infekcija (IMI) u suhostaju u programima suzbijanja mastitisa naglašava se značenje smještaja krava u suhostaju i prijelaznom razdoblju. Važnost kontrole IMI u suhostaju u svakodnevnoj praksi se često podcjenjuje., Mastitis is the most common disease in dairy cows and most antibiotics are used for treatment and prevention. Frequent, inappropriate, or unnecessary use of antimicrobials poses a major risk for accelerated development of antimicrobial resistance. Transmission of antimicrobial-resistant pathogens to humans is possible via milk and dairy products, and guidelines for the rational use of antimicrobials in veterinary medicine are prescribed. In dairy farming, most antibiotics are used to prevent mastitis at drying off. At the same time, antibiotics are used unnecessarily even in uninfected animals. An alternative strategy that promotes lower antimicrobial usage is selective dry cow therapy, where only cows with mastitis receive antibiotic treatment. However, an issue in the implementation of selective dry cow treatment is the criteria applied for selecting the cow for drying off with antibiotics. Cow selection protocols should be sufficiently accurate, easy to perform and interpret, cost-effective, safe, certifiable, and applicable in different areas of a country. The criteria for cow selection can be based on bacteriological culture of milk, somatic cell count at drying off, incidence of clinical mastitis in previous lactation, parity, or any of their combinations. Bacteriological culture of milk is the gold standard, though somatic cell count at the last milking has proven to be a reliable indicator for cow selection to drying off with antibiotics without negative effects on milk production. The thresholds of more...

The potentially negative effects of radiofrequency electromagnetic fields (RF-EMF) of higher frequencies have not been investigated on human and animal ejaculates so far. The aim of this study was to investigate the effect of RF-EMF at 3500 MHz on spermatozoa DNA integrity and vitality indicators in vitro exposed ejaculates of breeding boars. Ejaculate samples of 12 breeding boars aged from 1.5 to 3.5 years were obtained by manual fixation of the penis. Freshly diluted ejaculates of each boar were divided into a control (n = 12) and a test sample (n = 12). The test samples were exposed to RF-EMF at 3500 MHz and field strength of 10 V/m for 2 hours using gigahertz transverse electromagnetic chamber. In the both samples, exposed and non-exposed ejaculates, the proportion of spermatozoa with cell and acrosomal membrane damage, the proportion of spermatozoa with damage and depolarization of mitochondria using PI⁄SYBR- 14 LIVE⁄DEAD Sperm Viability Kit and the proportion of spermatozoa with DNA damage were determined using Halomax DNA Fragmetation Test. The significantly increase in the proportion of spermatozoa with DNA damage was recorded in ejaculates exposed to RF-EMF compared to non- exposed ejaculates. A significantly higher proportion of spermatozoa with damaged membrane and good mitochondrial potential was recorded in ejaculates exposed to RF-EMF at 3500 MHz. Although there was an increase in the sum of the proportion of spermatozoa with damaged membrane and/or low mitochondrial potential in all exposed ejaculates, but not significant. In vitro exposure of breading boars’ ejaculates to RF-EMF at 3500 MHz has a negative effect on the DNA integrity. Furthermore, the effect of RF-EMF at 3500 MHz on in vitro exposed ejaculates, is manifested by an increase in the proportion of spermatozoa with damaged membrane and/or low mitochondrial potential. more...

The aim of this study was to determine the influence of dietary clinoptilolite (CPL) on serum levels of Ca, Mg, P, K and Na of Holstein Friesian heifers (n=20) and cows (n=20) during late pregnancy and early lactation and their reproductive performance. They were randomly assigned into 2 groups (each containing 10 cows and 10 heifers): the CPL-treated group that received 100 g CPL in feed from 60 days ante partum (ap) to 40 days post partum (pp), and the control group (CON) of untreated animals. Blood samples were taken on days 30 ap, 10 ap, on the day of calving and on days 5, 19, 33, and 40 pp. There was a significant difference in Ca levels between the CPL and CON groups (2.41±0.16 vs. 2.28±0.12 mmol/L, respectively) 30 days ap. The Ca level was higher in the CPL group than in the CON group (2.60±0.14 vs. 2.19±0.13 mmol/L, respectively ; P more...

To the best of our knowledge, this is the first analysis of morphometric parameters for assessing the influence of radiofrequency radiation (RF) at 2500 MHz on spermatozoa subpopulations. The aim of this study was to establish the influence of RF at 2500 MHz in vitro exposed breading boars’ semen on proportions of spermatozoa subpopulations using the principal component (PC) and cluster analysis according to morphometric head and tail parameters. Semen of 12 breeding boars was collected using a manual fixation method. Semen samples of each boar were divided into a control and an experimental sample. The experimental samples were exposed to RF of 2500 MHz in gigahertz transverse electromagnetic chamber for 2 hours at an electromagnetic field strength of 10 V/m. Semen smears were stained using the Spermac method. Morphometric analyses in the control and experimental samples were performed in stained smears by SFORM computer- assisted program. Morphometric size and shape parameters of boar’s spermatozoa (n=1691) were determined. The PC analysis revealed four components with the most important value for each (head length and width, head regularity, midpiece outline). Cluster analysis indicated there were two subpopulations (S1 - spermatozoa of smaller length and larger head width, more regular head shape and smaller circumference of the midpiece and S2 - spermatozoa of longer length and smaller head width, more elongated head shape and larger outline of the midpiece). Exposed boars’ semen samples had a greater proportion of S1 spermatozoa compared to control samples (51.83% vs 48.7%) and a lesser proportion of S2 spermatozoa (47.54% vs 52.47%). The effect of RF at 2500 MHz in vitro exposed breading boars’ semen was manifested in a reduced proportion of spermatozoa subpopulation with the spermatozoa of more elongated head and larger midpiece outline which could decrease spermatozoa motility and fertility potential. more...

A 15-year-old intact female Pekingese was referred to the Clinic because of vaginal bleeding and polydipsia. The patient had a previous diagnosis of degenerative mitral valve disease ACVIM stage C, and was stable under standard therapy. An enlarged and painful abdomen was observed during examination. Haematology and biochemistry profile showed signs of inflammation, most probably due to infection, and increased BUN and creatinine levels. Abdominal ultrasound showed dilation of the uterus with anechoic area and right uterine horn filled with content. The patient was premedicated with midazolam (0.25 mg/kg IV) and methadone (0.15 mg/ kg IV). General anaesthesia was induced with propofol (3 mg/kg IV), followed by endotracheal intubation. The patient was connected to the anaesthetic machine with a rebreathing circuit and maintenance of the anaesthesia was provided by a combination of oxygen and isoflurane. Pain management was provided by the use of fentanyl boluses (2, 5 µg /kg IV). Patient monitoring included capnometry, non-invasive arterial blood pressure measurement, pulse oximetry, and ECG. Intraoperative findings were unilateral segmental aplasia of the left uterine horn and right horn filled with content and uterine body tumor. Ovariohisterectomy was performed with subsequent pathohistology. The pathohistology findings were: squamous metaplasia and cystic degeneration of the endometrial glands, uterine adenomyosis and squamous aplasia of the left uterine horn and left ovary. Postoperative care included pain management, antibiotic therapy, control of renal parameters and constant monitoring of breathing frequency with with attention to the occurrence of edema due to cardiac problems. The patient was discharged to home care two days after the procedure. In conclusion, this was a complex case of unilateral uterine horn aplasia, pyometra and uterine tumor complicated with a heart disease that in the end had positive outcome more...

With the development of technology, there is an increase in public concern about the negative effects of radiofrequency electromagnetic radiation (RF-EMR), especially of device that operates at a frequency of 2500 MHz. It is presumed that the male reproductive system could be one of the most affected by negative effects. The data on the effect of RF-EMZ on the reproductive system of male domestic animals are scarce. The aim of this study was to investigate the effect of electromagnetic radiation with a frequency of 2500 MHz on sperm motility and velocity parameters of in vitro irradiated semen of breeding boars. The ejaculates of 12 breeding boars, Landrace and Pietrain breed, aged 1.5 to 3.5 years were sampled by manual fixation of the penis. Freshly diluted semen of each boar were divided into a control (n = 12) and an experimental sample (n = 12) that was irradiated. The experimental samples were exposed to continuous non-ionizing electromagnetic radiation of 2500 MHz in gigahertz transverse electromagnetic chamber for 2 hours at an electromagnetic field strength of 10 V/m. In the control and experimental groups, the motility and sperm velocity parameters were determined using computer assisted sperm analyser. Following sperm velocity parameters were analysed: curvilinear velocity, straight-line velocity, average path velocity, amplitude of lateral head displacement, linearity, wobble, straightness and beat cross frequency. Significant decrease (p more...

The objective of this study was to evaluate the effects of dietary zeolite clinoptilolite on reproductive performance, serum progesterone and insulin-like growth factor 1 (IGF-1) concentrations in 78 Holstein Friesian (HF) cows during pregnancy and lactation. The cows were divided into two groups comprising 40 (control group ; CON) and 38 (CPL group) cows. To assess reproductive performance of HF cows the following variables were registered: the interval from calving to first insemination (days open to first service, DFS), the interval from calving to pregnancy (days open to pregnancy, DOP), and the number of services per pregnancy (NSP). The average values of progesterone (5.64±0.59 ng/mL vs. 5.16±0.64 ng/mL) were not statistically different (p0.05) were shorter in the CPL than in the CON group (115.1±19.9 and 137.5±36.3 days vs. 124.2±17.3 and 143.8±33.5 days, respectively). During 305 days of lactation, milk production was higher in the CPL vs. CON (8325.5±628.8 kg vs. 8050±586.8 kg). The NSP was lower in the CPL than in CON group (1.91 vs. 2.14). The dietary clinoptilolite supplement had a positive influence on milk yield, exhibited modulating effects on endocrine status of dairy cows, and improved reproductive performance, with the decreased NSP, and fewer DFS and DOP. more...

Follicular fluid (FF) proteins originate from blood plasma and theca, granulosa cells and oocyte. The FF proteins regulate follicular development and oocyte maturation. Among numerous functions, they are acute phase proteins (APP) and proteins with antioxidant properties. Ceruloplasmin and haptoglobin are APP and protect follicular cells from oxidative damage binding copper and haemoglobin. The aim of the study was to determine follicle size and estrous cycle phase related changes in concentration of ceruloplasmin and haptoglobin. For the better insight in non- protein antioxidative molecules, biological antioxidant potential (BAP) was determined. Oxidative processes intensity was examined by reactive oxygen metabolites (ROMs). The bovine ovaries were collected from naturally cycling animals at a local abattoir. The stage of estrous cycle (follicular or luteal) was identified according to presence or absence of the corpus luteum on the ovary. Follicular diameter was measured and follicles were classified according to diameter: small (≤5 mm ; N=18 luteal phase ; N=14 follicular phase), medium (6-10 mm ; N=14 luteal phase ; N=12 follicular phase) or large (>10 mm ; N=12 luteal phase ; N=14 follicular phase) follicles from ovaries in both follicular and luteal stage of estrous cycle. Ceruloplasmin and haptoglobin concentrations and biological antioxidant potential (BAP) and concentration of ROMs were analysed in FF punctured from ovaries in follicular and luteal phase of estrous cycle. Concentrations of BAP and haptoglobin significantly decrease from small to large follicles collected in both luteal and follicular phase. The highest concentration of ceruloplasmin in FF was determined in medium size follicles but changes was not statistically significant compare to small and large follicles. During folliculogenesis, in luteal phase follicles ROM concentration significantly decreased from small to large follicles. During folliculogenesis ceruloplasmin negatively correlate with haptoglobin but significant correlation was noted only in small follicles from luteal phase (r= - 0.70 ; p more...

In the original manuscript the affiliations for the authors M. Gerencer, A. Kastelan, A. Milovanovic, M. Lazarevic, D. Rukavina, I. Valpotic are set incorrectly. This has now been corrected as shown below. M. Samardzijaa , M. Lojkica , N. Macesica , H. Valpoticb , I. Butkovica , J. Savorica , I. Zura Zaja c , D. Leinerd , D. Duri - cice , F. Markovicf , P. Kocilag , Z. Vidas h , M. Gerencerl , A. Kastelani , A. Milovanovicj , M. Lazarevick , D. Rukavinai and I. Valpotic m a Clinic for Obstetrics and Reproduction of Animals, Veterinary Faculty University of Zagreb, Zagreb, Croatia; b Department for Animal Nutrition and Dietetics, Veterinary Faculty University of Zagreb, Zagreb, Croatia; c Department for Physiology and Radiobiology, Veterinary Faculty University of Zagreb, Zagreb, Croatia; d Department of Anatomy, Histology and Embriology, Veterinary Faculty University of Zagreb, Zagreb, Croatia; e Veterinary Practice, Durd - - evac, Croatia; f Belupo d.d. Danica, Koprivnica, Croatia; g Animal Feed Factory, Cakovec, Croatia; h Faculty of Medicine, Department of Urology, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia; i Croatian Academy of Sciences and Arts, Croatian Academy of Sciences and Arts, Zagreb, Croatia; j Department of Reproduction, Veterinary Scientific Institute, Novi Sad, Serbia; k Department for Physiology and Biochemistry, Faculty of Veterinary Medicine, University of Belgrade, Belgrade, Serbia; l Department of Cellular Immunology, Baxter Hyland Immuno, Vienna, Austria; mDepartment of Biology, Veterinary Faculty University of Zagreb, Zagreb, Croatia Link to the corrected article: [https://vet-erinar.vet.bg.ac.rs/handle/123456789/1967] more...

Background. Oxidative stress is important for promoting the oocyte maturation and ovulation within the follicle. The aim of the study was to examine activities of the superoxide dismutase (SOD), glutathione peroxidase (GPx) and biological antioxidant potential (BAP) involved in protection against free oxygen radicals and concentration of reactive oxygen metabolites (ROMs) in bovine follicular fluid. Methods. Bovine ovaries were obtained from a slaughterhouse. The stage of estrous cycle (follicular or luteal) was identified. Follicular fluids (FF) were collected by puncture from three categories of follicles: small (≤5 mm), medium (6-10 mm) or large (>10 mm) from ovaries in both follicular and luteal stage of estruos cycle. Results. The results indicate a significantly higher activity of SOD in bovine FF from follicles in luteal stage of estruos cycle than in follicular stage (p=0.037). In FF of follicles in both stages of estrous cycle, SOD activity and BAP were significantly higher in FF of small and medium sized follicles than in large ones (p=0.000 both). The ROMs significantly declined from small to large follicles in FF collected during both, luteal and follicular phase of estrous cycle (p=0.005 both). Activity of GPx showed no significant differences regarding neither estrous cycle nor size of follicles. Conclusions. Results indicate similar antioxidative properties of bovine follicles during both luteal/follicular stage of estrous cycle with higher SOD activity in FF of follicles in luteal stage of estrous cycles which undergo atresia. more...

The objectives of this study were to determine whether or not dietary clinoptilolite (CPL) has an influence on the levels of calcium (Ca), magnesium (Mg), phosphorus (P), potassium (K) and sodium (Na) in the blood serum of dairy cows during gravidity and early lactation. The study was conducted on 78 dairy cows of Holstein-Friesian breed. The cows were randomly assigned into two groups: the CPL- fed treated group (n=38) which received 50 g of natural powdered zeolite CPL twice a day from day 180 days before to 60 days after parturition, and the control non-treated group (n=40). Blood samples were taken on days 180, 90, 60, 30 and 10 before parturition, on the day of calving and on days 5, 12, 19, 26, 33, 40 and 60 following parturition. There were no significant differences in the Ca concentrations between the CPL-fed and the control group. However, after parturition it was noticeable that the Ca concentration was higher in the CPL-fed group, especially on day 33 (P=0.06). The concentration of P was significantly lower (P more...

Problemi vezani za reprodukciju i mastitise čine dva područja koja donose najveće gubitke u mliječnom govedarstvu. Stoga je neophodno pratiti najnovija istraživanja u vezi s prevencijom, otkrivanjem i liječenjem mastitisa. Kad god smo u mogućnosti treba u praksi provjeriti rezultate tih istraživanja jer nije sve primjenjivo u uvjetima koji vladaju na različitim farmama. Također treba provoditi vlastita istraživanja kad god je to moguće. Većina primijenjenih istraživanja kroz primjenu njihovih rezultata nudi smanjenje troškova proizvodnje i liječenja iz jednostavnog razloga. Gornja granica proizvodnosti životinja je praktički već odavno došla do svojih limita. Današnje krave iako genetskim potencijalom odgovaraju većoj proizvodnji mlijeka jednostavno ne mogu pojesti hrane tj. unijeti energije, dovoljno da bi se taj potencijal u potpunosti ostvario. Svako daljnje forsiranje visoke proizvodnje neminovno bi dovelo do ozbiljnih zdravstvenih problema. Sa sadašnjom visokom proizvodnjom to je ples po tankoj liniji između bolesti i zdravlja. Dovoljne su male promjene da životinje izgube tu ravnotežu i od zdravih postanu bolesne. Stoga je najvažniji zadatak veterinara konstantno praćenje zdravlja i proizvodnosti mliječnih krava (ili bilo kojih drugih proizvodnih životinja). Korištenje rezultata najnovijih istraživanja i upotreba novih generacija lijekova i vakcina pomoći će u održavanju stada zdravima uz visoku proizvodnju. more...

The relationship between the main characteristics of native semen and fertility in young military working dogs has been the subject of a very small number of studies. This research provides, for the first time in Croatia, a comprehensive semen evaluation of the military working dogs (MWD) owned by the Ministry of Defence of the Republic of Croatia (MDRC). Nine virgin MWD (5 German Shepherd dogs and 4 Belgian Shepherd dogs, Malinois), aged 12 to 18 months (1.24 ± 0.19 years) were subjected to a clinical examination, B-mode ultrasonography of the testes, epididymis and prostate, semen evaluation at collection and after storage at 4 °C for 24, 48 and 72 h. The semen was evaluated based on the following parameters: colour, volume, pH, progressive motility, concentration, viability and functional integrity of the sperm membrane (HOS test and Bloom staining). The diluted semen samples were subjected to a longevity exam at 4 °C, and the semen quality was assessed after 24, 48 and 72 hours. The dogs were subjected to the second semen collection and evaluation after two months. In 7/9 of the virgin dogs, the semen was successfully collected by using digital manipulation at the first collection and in 8/9 of the dogs at the second collection, without the presence of a teaser bitch. All sperm quality parameters were within the optimal range. Sperm progressive motility and motility after 24 and 48 hours of storage was significantly higher in the second collection (p < 0.05). The percentage of morphologically normal spermatozoa was also higher in the second-collection specimens, although without a significant difference. This research has shown that it is necessary to include a variety of clinical and laboratory methods in order to get a complete insight into the breeding potential of dogs and choose those with the highest- quality characteristics for reproduction. Our results of the semen quality evaluation at collection and after storage indicate a high reproductive potential of military breeding and working dogs of the MDRC and offer a possibility of international trade of a high- quality genetic material. more...

The idea of a dual-sired breeding is to give equal chances to both possible males to fertilise the eggs due to specific oestrus cycle of the bitch. Successful multiple-sired litters allow breeders to maximise offspring possibilities as well as the female’s potential by creating more genetic combinations. Three stud dogs (A, B and C) were presented to the clinic a month before breeding to undergo sperm quality testing in order to choose 2 potential sperm donors. All dogs had a history of prior normal fertility and had a DNA profile. The semen was collected by manual manipulation and only sperm-rich fractions were collected. Immediately after the collection, the semen was evaluated based on the following parameters: colour, volume, pH, progressive motility, concentration, viability and functional integrity of the sperm membrane (HOS test and Bloom staining). After the evaluation, the semen samples were mixed according to the factorial design Stud A and B, Stud A and C and Stud B and C. The mixed samples were evaluated for semen characteristics after 1, 2, 3 and 5 hours. In addition, the mixed samples were subjected to a longevity exam at 4 °C, and the semen quality was assessed on days 3 and 5. The breeding time of the bitch was determined by measuring progesterone (P4) levels. P4 concentration at the onset of ovulation was 4.2 ng/mL. The bitch was inseminated 3 times starting on day 2 after ovulation. Based on semen quality parameters, studs A and B were chosen for insemination. On all 3 days of insemination, the semen of 2 males was collected, evaluated and mixed prior to insemination. Semen volume was adjusted based on quality parameters, to give an equal final concentration for both males. The bitch was inseminated vaginally. The puppies were born 60 days after the first insemination. Blood samples were collected on week 5 via jugular venepuncture into EDTA vacutainer tubes for a genetic analysis. The final results showed 6 puppies from stud A (4 ♂, 2 ♀) and 1 puppy from stud B (♂). No monozygotic twins were born. more...

A 1-year-old Kangal Shepherd bitch was admitted to the Clinic due to ultrasound (US) diagnostic of the first pregnancy 30 days following mating. Canine pregnancy normally ranges from 58 to 70 days. The US finding revealed 4 foetuses with a physiological heartbeat. The gestational sac diameter corresponded to foetal age. A suggestion for the next control examination was at the 45th day of the gestation, but the owners came on the 91st day, since parturition had not started. The bitch underwent a physical examination, which was unremarkable. The CBC showed mild anaemia, which corresponds to mid gestation. Serum biochemistry results were in the normal ranges, except hypoalbuminemia and a slightly increased CRP. Serum P4 was 7.88 ng/mL. There were no vaginal signs for parturition initiation. An abdominal US showed well-developed but avital foetuses without foetal fluid. The endometrium was unchanged without intraluminal free fluid. The X-ray showed 6 completely developed and ossified foetuses without foetal fluid. There were also initial signs of skeleton decalcification. Based on the elevated P4 and the absence of the free fluid, mummification of the foetuses occurred due to persistence of corpus luteum presence. A radical C-section was performed in general inhalation anaesthesia and all the foetuses with placenta were delivered. The cytology of the smear and endometrium aspirate revealed mild cellular slide with a predominance of segmented neutrophils (86%), lymphocytes (12%) and eosinophils (2%) with haematoidin crystals, connected to non-septic neutrophil inflammation. The bacterial findings of uterine aspirate as well as herpes viral molecular testing of foetal organs were negative. The postpartum X-ray of foetuses showed no signs of congenital malformation. In conclusion, prolonged gestation can result in foetal death and many further complications. At the final stage of pregnancy, it is important to monitor the decline of serum P4 as the evidence of luteolysis in order to prevent a prolonged parturition and foetal death. more...

The aim of the study was to establish the influence of dietary clinoptilolite (CPL) supplementation on milk fat-to-protein ratio (FPR) and serum β-hydroxybutirate (BHB) levels during transition period in dairy cows. Thirty pregnant cows for 240 days of the Holstein- Frisian breed, aged from 3 to 5 years and kept in a farm near Đurđevac, Croatia, were used in the trial. The cows were randomly assigned into two groups (either in-feed CPL treated ; CPL group or control non-treated ; CON group) comprising 15 cows each. The CPL group received 100 g of natural modified clinoptilolite preparation (Vibrosorb, Viridisfarm, Podpićan, Croatia) in the ratio for dairy cows on a daily basis. Blood samples were collected from the cows in the period from day 240 to 260 of pregnancy, after parturition (day 0), and on days 12, 26, 40 and 60 of lactation. During lactation, milk samples were taken (same days when a blood samples were taken) for milk fat and milk protein analyses. The average value of BHB was higher in the CON than in the CPL group (0.99±0.10 vs. 0.81±0.99 mmol/L). The highest concentration of BHB was obtained on days 0, 12 and 26 of lactation in the CON group (1.83±0.35, 1.55±0.35, and 1.62±0.35 mmol/L, respectively) and in the CPL group after parturition, on day 0 (1.79±0.41 mmol/L). The FPR was significantly higher in the CON group on day 0, (P more...

Objectives: To study the effects of dietary zeolite CPL on P4 and IGF-1 blood concentrations in Holstein Friesian (HF) cows during pregnancy and early lactation. Methods: Twenty HF-cows, aged 3–5 years and kept on a commercial farm near Đurđevac, Croatia were used in the study. They were assigned into two groups, each of 10 cows. The cows from the treatment group received 100 g/day of natural CPL modified by vibroactivation and micronization (Vibrasorb, Viridisfarm, Podpićan, Croatia). Blood samples were taken on days 90, 180, 210 and 240 of pregnancy, on day 10 before and day 0 after parturition, and on days 5, 12, 19, 26, 40 and 60 of lactation. Serum concentrations of P4 and IGF-1 were determined using ELISA. The obtained data were statistically analysed using the ANOVA method with repeated measurements. Differences were considered significant at p < 0.05. Results: The average values of P4 (6.25 ± 0.73 ng/ml vs. 6.00 ± 0.58 ng/ml) and IGF-1levels (425.20 ± 59.36 ng/ml vs. 397.21 ± 41.23 ng/ml) were higher in the CPL-fed than in the control cows. During pregnancy, the highest level of P4 was recorded in the treatment group on day 90 (11.84 ± 1.78 ng/ml) and in the control group on day 240 (11.25 ± 1.69 ng/ml). In CPL-fed cows ovarian cyclity resumed on day 33 postpartum (PP), when the IGF-1 level was highest during the PP period, which was also higher (p < 0.05) than in the control cows (729.36 ± 190.39 ng/ml vs. 443.44 ± 141.91 ng/ml). Also, a consecutive increase of P4 levels was recorded from days 40 to 60 PP (5.84 ± 1.88 ng/ml and 7.87 ± 2.18 ng/ml). Conclusions: A dietary CPL preparation exhibited modulating effects on the endocrine status of dairy cows by increasing their P4 and IGF-1 serum levels during the PP period, which may influence their reproductive efficiency. more...

U ovom radu su opisani utjecaji clinoptiloita na koncentraciju progesterona i IGF-1 u krava holštajn-frizijske pasmine tijekom graviditeta i rane laktacije.

U veterinarskoj medicini sve se više operacija iz različitih kirurških grana izvode endoskopskim metodama, budući da su istraživanja ukazala na slabiji odgovor organizma na kiruršku traumu kod pacijenata operiranih endoskopskim načinom. Sukladno tome u kirurškoj reprodukciji sve veći broj operacija izvodi se endoskopskim načinom. Vjerojatno najčešće izvođena operacija u veterinarskoj medicini je kastracija kuje koja je i predstavljala početak minimalno invazivne kirurgije. Prva laparoskopska operacija u veterinarskoj medicini izvedena je davne 1985. godine i to je bila laparoskopska ligacija vrha roga maternice kuje. Prva laparoskopska ovarijektomija kuje izvedena je 2004. godine na Klinici za porodništvo i repredukciju Veterinarskog fakulteta Sveučilišta u Zagrebu. Od tada se na istoj Klinici rutinski izvode elektivne i medicinski indicirane laparoskopske operacije. Najčešće laparoskopski izvođeni zahvati na našoj klinici su: laparoskopske ovarijektomije i ovariotomie, laparoskopske ovarijohisterektomije, laparoskopske ovarijohisterektomije početnih stadija pijometre i cistične hiperplazije endometrija, laparoskopske intrauterine inseminacije, laparoskopske gastropeksije, laparoskopske kriptorhidektomije, laparoskopske operacije sindroma zaostalog jajnika i laparoskopske biopsije. Dokazano je da biološki odgovor na operaciju ovisi o jačini traume u odnosu na mogućnost oporavka životinje te je sasvim jasno da laparoskopska kirurgija nudi neke prednosti nad konvencionalnom kirurgijom. Laparoskopska ovarijektomija i ovarijohisterektomija mogu se izvoditi na više načina ovisno o dostupnoj opremi te uvježbanosti operatera. Nedostatak endoskopskih tehnika je u vrlo skupoj opremi te relativno zahtjevnoj i dugotrajnoj edukaciji operatera. Laparoskopske operacije uz sve prednosti imaju i kontraindikacije. Relativne kontraindikacije su ascites, abnormalno vrijeme grušanja krvi i eventualno pretilost. Apsolutne kontraindikacije za laparoskopiju su dijaframatska hernija. Komplikacije laparoskopskih operacija su slične kao u otvorenoj kirurgiji uz mogućnost nastanka termičkih ozljeda unutarnjih organa nastalih radom sa elektrokauterom koje nisu uobičajene u konvencionalnoj kirurgiji, a češće se viđaju u laparoskopskoj kirurgiji, kao i ozljede unutarnjih organa prilikom uvođenja trokara. Takve komplikacije mogu imati katastrofalne posljedice. Zbog toga, učenje laparoskopske kirurgije je nešto dulje i teže nego konvencionalne kirurgije. Prilikom laparoskopske operacije mora se koristiti endotrahealna inhalacijska anestezija s mehaničkom ventilacijom. Uspješnost laparoskopske kirurgije tj. mogućnost izvođenja laparoskopskih operacija usko je vezana uz dostupnost laparoskopske opreme, instrumentarija i puno je izraženija nego kod tradicionalne kirurgije. Postoji nekoliko metoda laparoskopske ovarijektomije: single incision laparoscopy, single port laparoscopy, operacija na dva paralelna porta, operacija na tri paralelna porta i operacija na tri porta pravilom triangulacije. Također, ovarijohisterektomija se može izvesti laparoskopski i manje zahtjevno laparoskopski asistiranom tehnikom. more...

Više od pet desetljeća istraživanja re- produktivne biologije omogućilo je razvoj biotehnoloških postupaka koji povećavaju učinkovitost mesnog i mliječnog govedar- stva. Ovi postupci omogućuju manipulaciju gametama i zametcima u svrhu poboljšanja plodnosti i genetskog napretka. Usvajanje i primjena postupaka asistirane reproduk- cije u rasplođivanju krava ima dalekosežne posljedice u komercijalnom uzgoju stoke. Umjetno osjemenjivanje, embriotransfer i proizvodnja zametaka in vitro tehnologije su koje se sustavno primjenjuju u uzgojnim pro- gramima diljem svijeta omogućavajući brzi genetski napredak, skraćenje generacijskog intervala, kontrolu zaraznih bolesti i sman- jenje proizvodnih troškova. Umjetno os- jemenjivanje desetljećima je najisplativiji i najkorišteniji način širenja poželjnog geno- ma. Embriotransfer je tijekom posljednjih 30 godina postao međunarodno prihvaćena metoda rasplođivanja s preko 500 000 zam- etaka proizvedenih in vivo godišnje. Propisi Međunarodnog društva za embriotransfer vezani uz sanitarnu kontrolu proizvodnje za- metaka omogućuju siguran transport in vivo proizvedenih zametaka, bez rizika od širenja zaraznih bolesti. Od ukupnog broja zametaka, oko 15% godišnje ih se proizvodi in vitro. Ova tehnologija omogućuje dobivanje zametaka od plotkinja visoke genetske vrijednosti koje ne mogu dati potomstvo prirodnim putem, a zbog veće učinkovitosti s vremenom bi mo- gla zamijeniti klasične programe MOET-a. Postupci asistirane reprodukcije poput em- briotransfera i in vitro proizvodnje zametaka zajedno s krioprezervacijom, od izuzetne su važnosti u očuvanju izvornih i ugroženih pasmina goveda. Njihova primjena nezaobi- lazna je u programima uspostave banke gena koji pomažu u očuvanju farmskih genetskih resursa. Osim navedenog, ove biotehnologi- je povećavaju ponudu visokokvalitetne stoke te primjenu naprednih biotehnologija. Određivanje spola spermija i zametaka nudi stočarima mogućnost odabira spola teleta, a time i isplativiju proizvodnju mesa i mli- jeka. Napredne biotehnologije poput kloni- ranja i transgeneze imaju veliki potencijal u rasplođivanju, no zbog troškova i loših rezultata one se uglavnom koriste u istraživanjima te u farmaceutskoj industriji., More than five decades of research in reproductive biology have resulting in the development of biotechnologies in the cattle industry to increase efficiency in beef and dairy production systems. These technologies are related to gamete and embryo manipulation aimed at improving fertility and genetic progress. The application of assisted reproductive technology in stockbreeding has tremendously altered the rate of genetic improvement in breeding programmes and strategies. Artificial insemination, embryo transfer and in vitro embryo production are technologies systematically applied in breeding programs around the world. They enable rapid genetic progress, shortening of the generation interval, control of disease transmission and reduction of production costs. Worldwide, artificial insemination has been the most efficient and useful way to improve the genetic quality of the herd. Over a period of thirty years, embryo transfer has become an internationally accepted technology with over 500,000 in vivo produced embryos per year. The recommended handling procedures of the International Embryo Transfer Society enable the safe export of in vivo derived embryos, without the risk of disease transmission. Approximately 15% of embryos produced annually are produced by in vitro technology. This technology enables embryo production from cows of high genetic merit that cannot produce offspring by conventional reproduction. Improvements in OPU/IVF programs would have a great impact on the cattle industry and could replace the traditional MOET programs in the near future. Furthermore, they are important for the development and operation of a gene bank for the cryoconservation of animal genetic resources, to preserve indigenous and endangered breeds of cattle. In addition to genetic progress, the application of these biotechnologies in animal breeding permits high quality breeding stock to be available on the market and enables the application of advanced technologies. Semen and embryo sexing allows for identification and selection of sex, which can assist in the more efficient management of resources. Cloning and transgenesis have great potential in the cattle industry, though due to their low efficiency and high costs, these technologies are predominantly applied in experimental settings and the production of pharmaceuticals. more...

Objectives. The aim of the present study was to investigate whether the supplementation of clinoptilolite (Vibrosorb®, Croatia) in the concentrate feed of dairy cows has an effect on udder health and chemical composition of milk in dairy cows over two consecutive years. Materials and methods. Forty-six clinically healthy Holstein-Frisian cows, aged between 3-5 years, were randomly assigned to one of two groups: control (n=23) and CPL-fed group (n=23). The CPL group received 50 g zeolite in the ratio for dairy cows, twice a day. Initial milk sampling was performed prior to adding CPL to feed. Additional sampling sessions were carried out on a monthly basis up to the 7th month of pregnancy, and next lactation starting from the first month of lactation and over the next 6 months. Milk samples were analysed for chemical composition (milk fat, proteins, lactose, solids non-fat dry and urea), somatic cell counts (SCC) and by bacteriological examination. The data were analysed using the program Stata 13.1 (Stata Corp., USA). The average values of milk components were compared by the Student t-test between two groups for each sampling point. In repeated samplings within the same group of cows, the values for a milk component were compared using the repeated ANOVA. Post hoc analysis of values was performed by the paired t-test. Results. The results showed that clinoptilolite supplementation had no adverse effects on the tested chemical composition of milk at any of sampling points. However, the chemical composition of milk was found to be more stable in CPL-fed cows, especially at next lactation. A statistically significant difference between single samplings in CPL-fed cows was found for milk fat and urea. The SCC in milk did not differ significantly between groups but was higher in the control group (412, 260 vs. 339, 980, respectively). Average milk fat, proteins, lactose, solids non-fat dry and urea were slightly higher in the control than in CLP-fed cows (4.38±0.77 vs. 4.02±0.56, 3.85±0.37 vs. 3.64±0.32, 4.38±0.16 v.s.4.37±0.14, 9.21±0.36 vs. 8.94±0.29, 21.66±7.00 vs. 20.90±5.11, respectively). The values recorded in the frequency of isolation of the mastitis causative agent differed significantly between groups (P=0.01). Cows from the control group had a four-fold higher and at next lactation had a seven-fold higher incidence of intramammary infections than CPL-fed cows. Conclusions. The observed differences in the content of a particular milk component and SCC between the groups of cows did not differ significantly at any of sampling points. The beneficial outcome of the study may be attributed to the antibacterial, immunostimulating, detoxifying and antioxidative effects of CPL on udder health. more...

Masti su u folikularnoj tekućini (FT)važan izvor energije za stanice, sudjeluju u izgradnji membrane stanica i mueđustaničnoj komunikaciji. Cilj rada bio je utvrditi sastav masnih kiselina FT tijekom razvoja folikula. Istraživanje je provedeno na jajnicima krava holštajnske pasmine uzetima na liniji klanja. FT je punktirana iz malih folikula veličine ≤5mm i folikula srednje veličine 5-8 mm. Lipidi su ekstrahirani iz FT i masnikoselinski sastav je analiziran plinskom kromatografijom. Najzastupljenije masne kiseline u FT su miristinska, linolna i oleinska kiselina. Tijekom folikulogeneze maseni udio miristinske kiseline i ukupnih zasićenih masnih kiselina značajno se povećao, dok se udio linolne, alfa- linolenske, eikozapentaenske , dokozapentaenske kiseline, ukupnih UFA, PUFA, ukupnih n-3 PUFA i omjer eikozapentaenske i dokozapentaenske kiseline značajno smanjio tijekom rasta folikula (p more...

Canid herpesvirus-1 (CaHV-1) is enzootic pathogen of dogs often related to reproductive disorders. Unfortunately, substantial data about risk factors influencing seroconversion and spreading of the infection are scarce. In period from 2014 to 2017 a total of 128 breeding bitches, with no history of vaccination, were randomly sampled from 24 different breeding kennels in Croatia. From each animal multiple sera samples, vaginal and nasal swabs were collected in different stages of reproductive cycle. CaHV-1 antibody titre was determined using virus neutralisation assay and possible viral excretion using PCR method described earlier. Investigated risk factors which could affect on level of CaHV-1 antibody titres were: age, estrous cycle, number of mating, kennel size, attendance at dog shows, attendance at hunt and hygiene. Statistical analyses were performed using Statistica v.13 (TIBCO Software Inc, 2017). Data (antibody titres) were assessed for significance using ANOVA. more...

This study determined the lipid profle and antioxidative properties of bovine follicular fluid (FF) and also investigated their possible changes during foliculogenesis and reproductive ageing. Ovaries of heifers and cows were collected at a slaughterhouse. Samples of FF were obtained from follicles of three sizes: 5 mm (N=40) and >10 mm (N=35). Concentrations of triacylglycerol (TAG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), phospholipids (PHL) and activities of glutathione peroxidase (GPx), superoxide dismutase (SOD) and paraoxonase 1(PON 1) were analysed. The concentrations of TC, LDL- C, HDL- C, PHL were signifcantly higher, while the activities of GSH-Px and SOD were signifcantly lower in FF of cows compared to heifers. During foliculogenesis in the follicles of the heifers, concentrations of TAG, NEFA and the activity of SOD signifcantly decreased. Furthermore, during foliculogenesis in the follicles of the cows, concentrations of LDL-C and TAG as well as the activity of SOD signifcantly decreased, whereas concentration of TC signifcantly increased. In conclusion, these fndings indicate that age, together with the growth and development of follicles, affects antioxidant enzyme activity and lipid concentrations. more...

In Gefangenschaft gehaltene Echsen leiden sehr oft unter Reproduktionsstörungen. Die häufigste Reproduktionspathologie bei weiblichen Echsen ist die postovulatorische Retention von Eiern (Dystokie). Dabei wird zwischen einer obstruktiven und einer nicht obstruktiven Dystokie unterschieden, also einer erschwerten (Oviposition) bzw. einer völligen Unfähigkeit zur Eiablage. Häufig sind diese Erkrankungen durch Haltungs- und Fütterungsfehler bedingt, weshalb Tierärzte über gute Kenntnisse auf diesem Gebiet verfügen sollten. Ursachen für die Dystokie können eine mechanische Obstruktion der Eileiter, Infektionen des Reproduktionstraktes, eine große Anzahl an Eiern, eine mangelhafte Kalzifizierung der Eischale oder auch die metabolische Knochenerkrankung (metabolic bone disease) sein. Die klinischen Anzeichen von Reproduktionsstörungen sind unspezifisch. Zu ihnen zählen Depression, Lethargie, Anorexie, Dyspnoe, Obstipation sowie eine Reaktionslosigkeit auf äußere Reize. Das unspezifische klinische Bild macht die diagnostische Aufarbeitung zu einer großen Herausforderung. Als erstes sollte durch Röntgen- und Ultraschalldiagnostik der Zölomhöhle bestimmt werden, ob es sich über präovulatorische oder postovulatorische Pathologie handelt. Tierärzte, die über gute Kenntnis der Ultraschall- und Röntgendiagnostik bei Reptilien verfügen, können die Anzahl und die Position der Eier sowie Eierschalenbildung (Kalzifizierungsgrad) abschätzen. Die Bestimmung hämatologischer und biochemischer Blutparameter sowie der Elektrolyte kann helfen, den klinischen Zustand des Tieres besser einzuschätzen. Auch eine Endoskopie in allgemeiner Anästhesie kann bei der Diagnosefindung behilflich sein.Es bestehen unterschiedliche therapeutische Möglichkeiten (manuelle Massage im Wasserbad, chirurgische Behandlung, hormonelles Behandlungsprotokoll mit Kalzium- und Oxytocinanwendung). Die Auswahl der Behandlungsstrategie hängt von unterschiedlichen Faktoren ab: Soll das Tier reproduktionsfähig bleiben? Wie lange dauert die Retention der Eier (Dystokie) bereits an? Sie hängt aber auch vom klinischen Zustand des Tieres, der Art der Reproduktionsstörung und Prognose nach der Behandlung ab. Je schneller die Behandlung der Dystokie eingeleitet wird, desto größer ist die Chance, die Reproduktionsfähigkeit zu erhalten. more...

Routine quantitative serum progesterone (P4) assay is the most common method to estimate the date of ovulation and consequently the whelping date, which is 63±1 days after ovulation (1, 2). Whelping is a result of hormonal changes, including a final drop in a P4 values, which sharply decreases from 4-5 ng/ml to below 2 ng/ml starting 24 hours before the onset of whelping (1). When P4 is validated on a daily basis, it is important to have in mind fluctuations in progesterone level during the day, depending on a rhythmic secretion of this hormone (3). Serum progesterone level was measured in total of 17 Labrador retriever bitches in a late pregnancy starting from 58 days post ovulation. Blood samples were taken daily, at the same time. Enzyme-linked fluorescent assay (ELFA) was used to estimate a serum progesterone level. Retrogradely, the data from last 72 hrs was taken. According to a different rate of decrease, values were divided into two groups: rapid decrease (range between P4 values in last 72 hrs were 4, 15±1, 03 ng/ml) and gradual (slow) decrease (range were 1, 54±0, 54 ng/ml). Rapid decrease in P4 values was recorded in 41.2% and gradual rate decrease in 58, 8% of the bitches. Variations in changes of P4 level can be related to the age of the bitches. In the group with rapid decrease (n=7) mean age was 2±0.65 yrs and in the group with gradual decrease (n=10) was 3, 3±1, 73 yrs. Primiparous bitches were 47, 1% of the all bitches and 62, 5% of them had a rapid drop in P4 values. Consequently, percentage of the primiparous bitches in a group with a rapid decrease were 71.4% and in a group with gradual decrease were 30%. The size of a litter and the number of healthy puppies per litter doesn't seem to affect the rate of P4 decrease. Both groups had similar total number of the puppies (6, 6±2 and 7, 2±3 puppies for rapid and gradual P4 decrease) and the number of the healthy puppies (6, 1±1, 5 and 5, 1±3, 1 for rapid and gradual decrease). However, the number of the dead puppies per litter could be related to a rate of P4 decrease. Of all the newborn puppies 20, 3% were dead, 6, 5% in rapid decrease and 29, 2% gradual decrease group, respectively. Dystocia was present in 35, 3% of the all bitches, 28, 6% in group with rapid decrease and in 40% in group with gradual decrease, respectively. According to these findings, a rate of prepartum decrease in P4 level can differ greatly between bitches, which disables prediction of a precise time of whelping. Rapid decrease of P4 level can be expected in a young primiparous bitches. This quick hormonal turnover can be related to the stress of the first whelping, fast metabolism in a young animal and also can be a predictor of the healthy litter. more...

Capercaillie (Tetrao urogallus L.) is endangered bird species from the group of forest grouses (Tetraoninae). Despite still being present in the majority of its previous habitats, the population number is rapidly declining. Therefore, measures aimed at habitat analysis, understanding of factors with negative influence and establishment of artificial breeding are very important in preserving this species. Capercaillie inhabits mountainous areas, of Lika, Gorski kotar and Velebit, above 1000 m.a.s.l. The mating season begins at the end of the March and early April, with environmental temperature reaching 8 to 17 ° C. Out of sixty days of display, actual mating lasts for approximately only 8 days. Furthermore, in the artificial breeding, problems like failing to copulate, aggressive behaviour of males and females, avoiding of certain males, etc., were observed. The study was carried out in capercaillie breeding centre Prezid. The centre consists of 2 males and 5 females. At the time of the study they were at the end of the mating season, but without a successful mating. We took the sperm from the males by the lumbar massage method in the Eppendorf tube. Immediately after ejaculation, the sperm was diluted with a diluent (ASG medium) in a ratio of 1:2. Prepared diluted sperm was administered through the cloaca to the initial part of the uterus by a pipette. On the fifth day after artificial insemination, one of the eggs was cracked during the laying and analysed. On the surface of the egg yolk, we identified the embryo plate, and confirmed successful fertilization. Artificial insemination is a spare method in the case when natural fertilization fails, but also in the case of delayed egg production by females when males are already non-responsive. more...

Lipids are an important component of the follicular fluid (FF) as a cellular energy source and also have important biological functions in cell membrane biogenesis and signalling. Fatty acids (FAs) in FF are present in esterifed form as triacylglycerol (TAG), cholesterol esters and phospholipids (PHL), and as nonesterifed FAs (NEFAs). The aim of the study was to determine the lipid composition of bovine FF during folliculogenesis. Bovine ovaries were collected at a slaughterhouse, and follicular fluid samples were obtained from follicles of three different sizes (5 mm N=22 ; and >10 mm N=23). Concentrations of TAG, PHL, total cholesterol (TC), low- density lipoprotein cholesterol (LDL- C), high- density lipoprotein cholesterol (HDL- C), NEFA and FAs in FF were analysed. The saturated fatty acids were dominant in bovine FF and the most common was myristic acid (C14:0). Polyunsaturated fatty acids were the second most represented, with linoleic acid (C18:2) in the highest percentage. The least represented were monounsaturated fatty acids (MUFA), with oleic acid (C18:1c9) as the most common. The percentage of C14:1, C15, C18:1c9, C20:5n-3, C22:5, MUFA, and concentrations of LDL-C, TAG and NEFA were signifcantly higher in follicles 10 mm. The results also show the signifcantly lower percentage of C18:0, the ratio of arachidonic acid/eicosapentaenoic acid (C20:4n-6/C22:5n-3) and TC and PHL concentrations in follicles sized < 5 mm compared to follicles more than 10 mm in diameter. Our data show that differences in the lipid composition of FF are associated with the stage of follicle growth and development. more...

Postovulatory egg retention (egg binding/dystocia) is one of the most common reproductive disorders in captive female lizards, resulting in difficulties or complete inability of oviposition. Mechanical obstruction of the oviduct, large number of eggs or insufficient calcification of eggshell and Metabolic Bone Disease are the most common causes of egg retention in reptiles. A 2 year- old bearded dragon (Pogona vitticeps), weighing 300g was admitted to the Faculty Clinic due to interrupted oviposition. Four days before admission, the lizard layed one egg with an insufficiently calcified eggshell. From that time the lizard had been lethargic, without appetite and had been digging holes in substrate of terrarium. The animal is kept in adequate conditions. The owners did not add calcium supplement to its diet, so that may be a possible reason for unsuccessful oviposition. During clinical examination, hard consistency formations were palpable on ventral side of the distended celom. The lizard did not respond to pain during palpation. Ultrasound and X - ray findings confirmed the presence of 14 calcified eggs in the caudal region of the celom. There was no presence of free fluid in the coelomic cavity. Due to the fact that the eggs were well calcified, the diagnosis suggested post- ovulatory dystocia. Therapy included rehydration and stabilization of the patient (warm Ringer lactat+ 5% glucose 2% body weight s/c) and induction of oviposition. Calcium borogluconate at a dose of 35 mg/kg was administrated i/m. After 30 minutes, a second dose of Ca-borogluconate (50mg/ kg i/m) was administrated and 5 IU of oxytocin was given i/m. The animal was released from the Clinic to proceed with oviposition in her own terrarium. Within the next 20 hours the patient laid 14 calcified eggs. On X-ray follow up 24 hours after therapy, no presence of retained eggs were observed. The owners were advised that the current diet should be supplemented with calcium in the prescribed daily dose, to prevent future dystocia. more...

Jedan od glavnih čimbenika loše kvalitete sjemena, a time i reproduktivne funkcije muških životinja, upra vo je oksidacijski stres. To se stanje razvija bilo da se radi o povećanom stvaranju kisikovih i dušikovih reaktivnih spojeva (ROS/RNS) bilo smanjenoj mogućnosti antioksidacijske obrane zbog smanjene sinte ze ili unosa antioksidacijskih spojeva kao što su vitamin E, A, C, karotenoidi i selen. Membrana spermija-- sisavaca bogata je višestruko nezasićenim masnim kiselinama koje su osjetljive na peroksidaciju uzrokovanu reaktivnim kisikovim spojevima. Zaštita spermija od oksidacijskog stresa od najveće je važnosti. Spermiji i sjemena plazma sadržavaju enzime superoksid- dismutazu, glutaton- peroksidazu, katalazu, paraoksonazu 1, glutation, albumine i vitamine E i C. Sjemena plazma ima veću koncentraciju antioksidansa od drugih bioloških tekućina te nadoknađuje manju razinu ovih molekula prisutnih u spermijima. Glavni izvor reaktivnih kisikovih spojeva jesu nezreli i mrtvi spermiji kao i leukociti koji kontaminiraju sjeme. more...

Uzroci pobačaja u kuja i uginuća štenadi u ranoj dobi uglavnom ostaju nepoznati. Jedan od mogućih uzroka je pseći herpesvirus-1 (eng. Canine herpesvirus-1, CHV-1). Pretpostavlja se da se virus enzootski pojavljuje u cijelom svijetu i istraživanja u nekoliko europskih zemalja ukazuju na visoku seroprevalenciju u pasa. U ovom trenutku, prisutnost CHV-1 u Republici Hrvatskoj (RH) nije istražena niti je rađena poveznica s reproduktivnim problemima u kuja. Ukupno sedam štenaca iz tri različita legla (tri pobačena, dva mrtvorođena i dva uginula u dobi od tri i četiri dana) dostavljena su na analizu u Virološki laboratorij Zavoda za mikrobiologiju i zarazne bolesti s klinikom Veterinarskog fakulteta Sveučilišta u Zagrebu. Od unutarnjih organa, za analizu metodom lančane reakcije polimeraze (engl. Polymerase Chain Reaction - PCR), izdvojena je ukupna DNK iz jetre, slezene i pluća uginule štenadi te plodnih voda kuja. Metodom PCR-a dokazana je prisutnost CHV-1 virusa u jetrama navedene štenadi. Ovo istraživanje ukazuje na prisutstvo CHV-1 u kuja sa reproduktivnim poremetanjama u RH. Također, izolacija CHV-1 iz jetara uginule štenati može ukazivati na povezanost CHV-1 i pobačaja u zadnjoj trećini graviditeta, mrtvorodstvom te uginućem štenadi u prva četiri dana života. more...

The aim of the present study was to investigate the effect of follicle size from which oocytes originate on their subsequent ability to be fertilized and to undergo early embryonic development in vitro. Cumulus oocytes complexes (COCs) aspirated from small (≤ 5 mm) and large (> 5 mm) follicles of abattoir-derived ovaries were graded for their morphological appearance and were cultured to assess their developmental competence. In order to study the kinetics of early cleavage, the number of cleaved embryos was recorded at 24, 27, 30, 33, 36 and 48 hours post insemination (hpi). A rapid rise in the rate of cleaved embryos was observed by 27 hpi, for both oocytes collected from ≤5 mm and >5 mm follicles. However, oocytes recovered from follicles >5 mm cleaved in significantly higher proportion at 24 to 30 hpi, which was reflected to higher overall cleavage rate at 48 hpi. The kinetics of early cleavage was consistent with subsequent development of embryos, that is, the oocytes from >5 mm follicles which completed the first cell division faster developed to morula stage at Day 5 and blastocyst stage at Days 6 and 7 in higher proportion than oocytes from ≤5 mm follicles. Hatching rate on Day 9 was significantly higher when oocytes originate from follicles > 5 mm. Total cell number was not affected by follicle size. The results showed that oocytes derived from follicles > 5 mm display higher developmental competence than oocytes from ≤5 mm follicles, in terms of timing of first cleavage, timing of blastocyst development and overall blastocyst rate. The selection of oocytes based on follicle size and kinetics of cleavage could be useful tools in selecting the best embryos for transfer. more...

The objective of this research was to evaluate the Zagreb mastitis test (ZMT) as screening test for mastitis diagnosis in dairy cows. The study was carried out within the framework of subclinical mastitis monitoring in the Varaždin region. Research included 1549 quarter milk samples from 389 Simmental dairy cows. Milk samples for bacteriological examination, mastitis testing and SCC were taken in a sterile plastic tube during milking. Bacteriology examination of milk samples obtained 760 (49.06%) positive samples. The most common isolated pathogens, Staphylococcus aureus, Streptococcus spp. Lancefield group D, Pseudomonas aeruginosa, were found in 388 (51.05%), 84 (11.05%), 62 (8.16%) positive quarter samples, respectively. Using a cut-off of ZMT by scores ≥ 1 as positive, 63.20% (CI 95%, 60.80% - 65.60%) quarter milk samples were estimated as being positive for subclinical mastitis. This cut-off had quite high sensitivity (96.84%, CI 95%, 95.60% - 98.06%) and negative predictive value (95.79%, CI 95%, 94.14% - 97.44%), however specificity (69.20%, CI 95%, 65.98% - 72.74%) and positive predictive value (75.17%, CI 95%, 72.47% - 77.86%) were low. Using a ZMT cut-off ≥4×105 cell/mL as positive, 49.56% of the quarters were positive. Sensitivity (88.37%, CI 95%, 86.06% - 90.68%) and specificity (86.98%, CI 95%, 84.57% - 89.38%), as well as predictive values, were similar. On the basis of the results of this study, comparing SCC and ZMT in mastitis diagnosis, ZMT is a reliable diagnostic method for use in field conditions. more...