Tomasz Stankiewicz

Plasma progesterone analysis by a time-resolved fluorescent antibody test to monitor estrous cycles in goats

The objective of the current study was to evaluate whether blood plasma progesterone (P4) measurements with a time-resolved fluorescent antibody test (TR-FAT) kit designed for humans was applicable for goats. The first experiment was designed to verify whether the concentrations of P4 measured by TR-FAT can be used to monitor the estrous and ovarian activity in goats (n = 14). Blood samples (322) were collected, and the ovaries were scanned using ultrasonography. The second experiment was carried out on 4 goats (60 samples) and designed to compare the TR-FAT with radioimmunoassay (RIA). The time interval between the lowest concentrations of P4 assayed by TR-FAT was 21 ± 0.3 days and did not differ significantly from the length of the interestrous interval. The highest concentrations of P4 were confirmed by detection of corpus luteum. During estrus, the mean concentration did not differ significantly between both methods. Significant differences were present during the luteal phases; however, the profiles of P4 assayed by both methods followed a similar pattern. Regression analysis showed a correlation between the 2 methods (r = 0.98; r 2 = 0.96; P < 0.0001). The Bland-Altman plot showed that all averages were within the 95% limits of agreement; however, the differences between both methods tend to be greater as the average increases. The results demonstrated that the TR-FAT method can be applied to monitor estrous cycles in goats through measurements of plasma P4 concentrations. Moreover, not only does the TR-FAT meet the requirements for safety, but it is also a method of high throughput, rapidity, and simplicity.

Bacterial Contamination of Boar Semen and its Relationship to Sperm Quality Preserved in Commercial Extender Containing Gentamicin Sulfate.

This study was designed to determine the degree and type of bacterial contamination in boar semen (79 ejaculates from Large White and Landrace boars) and its consequences for sperm quality during storage (27 extended semen samples, 16°C for five days) under practical conditions of artificial insemination (AI). The results revealed the presence of aerobic bacteria in 99% of the ejaculates (from 80 to 370 ×106 colony-forming units/mL). Most of the ejaculates contained two or three bacterial contaminants, while the Staphylococcus, Streptococcus, and Pseudomonas bacterial genera were most frequently isolated. Also detected were Enterobacter spp., Bacillus spp., Proteus spp., Escherichia coli, P. fluorescens, and P. aeruginosa. In general, the growth of certain bacterial types isolated prior to semen processing (Enterobacter spp., E. coli, P. fluorescens, and P. aeruginosa) was not discovered on different days of storage, but fluctuations (with a tendency towards increases) were found in the frequencies of Bacillus spp., Pseudomonas spp., and Staphylococcus spp. isolates up to the end of storage. Semen preserved for five days exhibited decreases in sperm motility and increases in the average number of total aerobic bacteria; this was associated with sperm agglutination, plasma membrane disruption, and acrosome damage. We inferred that, due to the different degrees and types of bacterial contaminants in the boar ejaculates, the inhibitory activity of some antimicrobial agents used in swine extenders (such as gentamicin sulfate) may be limited. Because such agents can contribute to the overgrowth of certain aerobic bacteria and a reduction in the quality of stored semen, procedures with high standards of hygiene and microbiological control should be used when processing boar semen.

Concentrations of bone morphogenetic protein-15 (BMP-15) and growth differentiation factor-9 (GDF-9) in follicular cysts, mono - and polyoocyte follicles in gilts

Abstract The objective of the study was to determine the concentration of BMP-15 and GDF-9 in the fluid of follicular cysts and ovarian follicles, and to compare their concentrations in mono- and polyoocyte follicles in gilts. The study involved two experiments conducted on the ovaries collected post-slaughter from gilts (7-8 months old). The first experiment covered 31 follicular single cyst gilts (15-25 mm in diameter) and 41 gilts without cysts. Follicular fluid from follicles of 8-10 mm in diameter (n=41) and 5-8 mm in diameter (n=41), and cystic fluid (n=31) were collected for analysis. The second experiment involved collecting follicular fluid from poly- (n=19) and monooocyte (n=22) follicles. The concentration of BMP-15 and GDF-9 was then determined in the samples using specimen-specific ELISA kits. The differences in the concentration of these factors were calculated by means of analysis of variance and a posthoc test. Duncan’s multiple range test was used to verify the significance of differences at P<0.05 and P<0.01. In addition, correlations between the factors were calculated. BMP-15 and GDF-9 levels in the cystic fluid were significantly higher than those in the follicular fluid (P<0.01). However, no differences were observed between various size follicles or between mono- and polyoocyte follicles. BMP-15 and GDF-9 concentrations were found to be positively correlated (P<0.01). Differences in BMP-15 and GDF-9 concentrations in ovarian follicles and follicular cysts, as evidenced by our study, indicate that these factors may be related to folliculogenesis disorders in gilts. What is more, the number of oocytes in ovarian follicles does not influence the intrafollicular concentration of BMP-15 and GDF-9.

Relationship between the concentration of bone morphogenetic protein-15 (BMP-15) and growth differentiation factor-9 (GDF-9) in pre-ovulatory follicles, ovarian cysts and serum in sows

The aim of the study was to compare and assess the relationship between peripheral and ovarian bone morphogenetic protein-15 (BMP-15) and growth differentiation factor-9 (GDF-9) concentration in cysts-bearing and non-cysts-bearing sows. The research involved multiparous sows (cysts-bearing sows, n = 20; non-cysts-bearing sows, n = 26). Serum, cystic follicular fluid or fluid from the ovulatory follicles was collected from each of the sows at the time of the slaughter. The collected serum and cystic/follicular fluid samples were tested for BMP-15 and GDF-9 concentrations. BMP-15 and GDF-9 concentrations in fluid from ovarian cysts were significantly higher than in the follicular fluid (P < 0.01). The concentration of BMP-15 and GDF-9 in the serum of sows with cysts was also higher than in ovulating sows (P < 0.05 and P < 0.01, respectively). The concentration of BMP-15 in serum was positively correlated with the concentration of said protein in pre-ovulatory follicles and cysts (r = 0.70; P < 0.01 and r = 0.50; P < 0.05, respectively). A positive correlation was also found between the concentration of serum GDF-9 and cyst GDF-9 (r = 0.55; P < 0.05). Moreover, another positive correlation was detected between the BMP-15 and GDF-9 concentrations, with the highest correlation coefficient recorded in the cyst fluid (r = 0.87; P < 0.01). This study for the first time demonstrates the presence of BMP-15 and GDF-9 in the cyst fluid in sows and determines a positive correlation between the concentration of the above factors in the follicular/cystic fluid and serum. Therefore, it is possible that the determination of BMP-15 and GDF-9 serum concentration may assist diagnosing ovarian cysts in sows. However, it is suggested that further studies involving a more in-depth analysis of the role of BMP-15 and GDF-9 in the pathogenesis of ovarian cysts in pigs are needed.

Circannual changes in serum concentrations of thyroxine, calcitonin and parathormone in immature and mature red deer females (Cervus elaphus).

The aim of this study was to determine circannual changes in the serum concentrations of thyroxine, calcitonin and parathormone in mature and immature red deer females. Blood samples from 8 hinds were collected monthly for 26 months. Secretions of thyroxine and calcitonin showed circannual rhythms with significantly higher levels in the immature hinds compared to the mature animals (p<0.05). For thyroxine, the concentration was higher in the winter/spring period than in summer/autumn (p<0.05), while for calcitonin the concentration profile was the opposite (p<0.05). The concentration of parathormone was significantly higher in summer/autumn that in the other months of the experiment (p<0.01). These results may indicate that the hormones investigated may be involved in the regulation of seasonal reproductive activity and in processes contributing to entering puberty in red deer females.

Macro-elements composition of cystic and follicular fluid in the ovaries and their relationship to peripheral blood concentration in sows.

Abstract The aim of this work is the comparison of sodium, calcium, magnesium and phosphorus concentration in pre-ovulatory follicles and follicular cysts and their concentration in the serum of sows. The research was conducted on sows (Polish Large White x Polish Landrace crossbred). Sodium concentration in cystic fluid was higher than in the follicular fluid (P<0.05). Sodium concentration in the serum of cysts-bearing sows was higher than in non-cysts-bearing sows (P<0.01). Differences were also observed between cysts and serum of cysts-bearing sows (P<0.01), as well as follicles and serum of non-cysts-bearing sows (P<0.01). Calcium concentration in the serum of non-cystsbearing sows was higher than in the case of cysts-bearing sows (P<0.01). Differences were also between calcium concentration in the cysts and its concentration in the serum of cysts-bearing sows (P<0.05). Magnesium concentration in the serum of cysts-bearing sows was lower than in non-cysts-bearing sows (P<0.01). Differences in magnesium concentration were also between cysts and serum of cysts-bearing sows (P<0.01), as well as between follicles and serum of non-cysts-bearing sows (P<0.05). Phosphorus concentration in the cysts was lower than in the follicles (P<0.05). It was also lower in the serum of non-cysts-bearing sows than in the follicles (P<0.05). The concentration of sodium, magnesium and phosphorus in the serum was positively correlated with the concentration of the mentioned macro-elements in ovarian structures (r = 0.66, P<0.01; r = 0.49, and r = 0.48, P<0.05, respectively). A negative correlation was found between the concentration of serum calcium and the ovarian structures (r = -0.47, P<0.05). The differences in the concentrations of macro-elements suggest that the ionic composition may be one of the factors associated with the processes occurring in the ovarian structures of pigs. Cilj ovog rada je poređenje koncentracije natrijuma, kalcijuma, magnezijuma i fosfora u pre - ovulatornim folikulima i folikularnim cistama i njihove koncentracije u krvnom serumu krmača. Istraživanje je sprovedeno na krmačama (poljska velika bela x poljski landras). Koncentracija natrijuma u cističnoj tečnosti bila je već a nego u folikularnoj tečnosti (p<0.05). Koncentracija natrijuma u serumu krmača sa ovarijalnim cistama bila je već a nego kod krmača koje nisu imale ciste (p<0,01). Razlike su takođe primećene između koncentracija Na u sadržaju cisti i krvnom serumu krmača bez cisti (p<0,01), kao i između koncentracija u sadržaju folikula i seruma krmača bez cisti. Koncentracije kalcijuma u serumu krmača bez cisti bio je već i nego kod krmača sa cistama (p<0,01). Takođe su uočene razlike između koncentracije kalcijuma u cistama i njegove koncentracije u serumu krmača sa ovarijalnim cistama (p<0,05). Koncentracija magnezijuma u serumu krmača sa ovarijalnim cistama bila je niža nego kod nego kod krmača kod kojih nije zabeleženo prisustvo cista na jajnicima (p<0,01). Takođe su zabeležene razlike u koncentraciji magnezijuma u sadržaju cista i u serumu krmača sa cistama (p<0,01), kao i između koncentracije Mg u sadržaju folikula i seruma krmača bez cisti (p<0,05). Koncentracija fosfora u cistama bila je niža nego u folikulima (p<0,05). Takodje, koncentracija fosfora je bila niza u serumu krmača bez cisti (p<0,05). Koncentracija natrijuma, magnezijuma i fosfora u serumu bila je u pozitivnoj korelaciji sa koncentracijom navedenih makro- elemenata u strukturama jajnika (r = 0,66, p<0,01; r = 0,49; r = 0,48, p<0,05). Ustanovljena je negativna korelacija između koncentracije kalcijuma u serumu i u strukturi jajnika (R = - 0,47, p<0,05). Razlike u koncentracijama makro elemenata sugerišu da jonski sastav može biti jedan od faktora koji su povezani sa procesima koji nastaju u strukturama jajnika svinja.

The effect of chitosan on the concentration of 17β-estradiol and free triiodothyronine in mice exposed to polychlorinated biphenyls (PCBs).

The aim of this work was to examine (i) how the applied PCB mixture influences the level of 17β-estradiol (E2) and free triiodothyronine (FT3) in the blood plasma of mice (C57/BL/6J) and (ii) whether supplementation with chitosan would protect against the observed changes in the examined plasma hormone concentrations. In the study we used a mixture of indicator PCBs (CB no. 28, 52, 101, 118, 138, 153, 180) and our results showed their anti-estrogenic effects. Exposure to the mixture resulted in a significant decrease (P < 0.05) in plasma concentration of E2 relative to the control, and chitosan administration did not prevent the decrease. To the contrary, E2 concentration in the blood plasma of the mice which received both the PCB mixture and chitosan was lower compared to those which did not receive chitosan. Exposure to the PCBs also resulted in a decrease in FT3 concentration in the treatedgroup, although it was not as pronounced as for E2 and was prevented with dietary supplementation with chitosan, with the observed FT3 level in the chitosan-treated group similar to the control. In summary, supplementation with chitosan can only to a certain extent minimize the negative effects of exposure to PCBs.