IgG,IgA, and lysozyme in Martina Franca donkey jennies and their foals

Because immune transfer from jenny to donkey foal is mostly unknown, the aim of the present study was to evaluate, from 5 days before to 10 days after foaling, immunoglobulin (Ig)G, IgA, and lysozyme peripartal concentrations in serum and mammary secretions of 10 healthy, spontaneously foaling Martina Franca jennies and in serum of their mature, viable, healthy foals, in the first 10 days after birth. The results showed that, in jennies, mammary secretion of IgG levels (ranging between 16 and 75 mg/mL) and IgA (0.9–2 mg/mL), and IgG (6.8–13.5 mg/mL) and IgA (0.5–2.4 mg/mL) serum concentrations were not different along the time of study. Also, IgG concentrations in serum of foals did not show significant differences although a high level was observed at 12 hours after birth (8 mg/mL), and IgA concentrations in serum of foals did not show any significant difference, although a high level was observed at 12 hours after birth (1.2 mg/mL). Lysozyme increased significantly at Day 2 after parturition in mammary secretions of jennies (551.9 μg/mL) and at 12 hours in serum of foals (25.9 μg/mL). The study demonstrated that the pattern of passive immune transfer in donkey foals seems to be similar to that reported for the horse foal, with IgG predominating IgA in serum and mammary secretions of the jenny and also in serum of foals. The most significant early increase in foals' serum concerns lysozyme, which probably plays an important role in the innate immunity of the donkey foal in the first challenging hours after birth.     

Characteristics of ovarian follicular dynamics throughout the estrous cycle of Egyptian buffaloes

Data of 56 normal and 9 abnormal estrous cycles were collected from 9 Egyptian buffaloes (Bublus bublis) to describe the follicular growth wave pattern. Heat was checked twice daily while, ovaries were scanned daily to monitor the patterns of follicular waves. Day of ovulation was determined when the largest follicle was replaced by corpus haemorrhgicum (CH). Number of waves/cycle, day of emergence of the follicular wave, characteristics of the dominant follicle and corpus luteum (CL) growth features were monitored. Buffaloes displayed mainly two types of follicular waves; two (46.4%) and three (53.6%). In cycles of three wave pattern, time of emergence of the 1st wave post-heat was longer (P < 0.05) and number of recruited follicles/wave were larger (P < 0.05) compared to the corresponding values of the two wave pattern. Number of recruited follicles in early follicular waves (1st or the 2nd) had larger number (P < 0.05) compared to the subsequent ones. Follicles that reached ovulation in both types of estrous cycle had shorter life-span (P < 0.05) than the previous ones. Life-span of CH, growing and regressed CL were 3.6 ± 0.6, 11.2 ± 0.8 and 4.4 ± 0.5 days, respectively with no difference in both types of follicular wave. Three types of ovarian disorders were observed. Follicular waves and CL growth features showed unique pattern for each individual. These results demonstrate that buffaloes display two main types of follicular waves with dominance of three wave type.     

Morphological features of the placenta at term in the Martina Franca donkey

This study was designed to establish the morphological features of the placenta of the Martina Franca jenny. Ten placentas were harvested at the time of foal delivery and examined both for gross and histological characteristics. The following factors were determined: the total weight and volume of the placenta and its components, the surface area of the allantochorion, umbilical cord length and site of insertion, and the diameter of the umbilical cord vessels and vascular pattern type. The weight of the placenta was similar to previously reported for ponies, and represented 12% of foal birth weight. Umbilical cord length was similar to that in the horse but longer than in the pony, while cord weight was intermediate between the two. In a histological examination, numerous strong villi were observed at sites corresponding to the non-pregnant and pregnant horn and uterine body. No villi were detected in the area overlying the cervical star. Despite obvious similarities between the donkey and horse placenta, specific morphological features do exist, and are possibly related to the differences in length of gestation.     

Apoptosis in the canine endometrium during the estrous cycle

Apoptotic cell death in the endometria of 58 female dogs in different stages of the estrous cycle was assessed (in formalin-fixed, paraffin-embedded sections) with both the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay and immunohistochemical detection of caspase-3 activity. For both techniques, the apoptotic index was determined in the surface epithelium, stroma, crypts, and basal glands by counting the percentage of stained cells in a total of 500 cells in each category. In the surface epithelium and stroma, TUNEL- and caspase-3-positive cells were rare (apoptotic index<1) throughout the estrous cycle. However, caspase-3 detection showed a significant increase in the apoptotic index in the stroma during anestrus as well as an increase in the index in both the stroma and surface epithelium in late metestrus. The apoptotic index increased during late metestrus and anestrus in the crypts and basal glands; in the crypts, this increase was significant only when caspase-3 detection was used, whereas in basal glands, significant differences were found for both techniques. In conclusion, apoptosis was present in canine endometrial cells during the estrous cycle, but caspase-3 detection showed more significant differences than the TUNEL assay. Furthermore, a high apoptotic index (suggestive of endometrial desquamation) was not detected in the surface epithelium and there was no significant correlation between the apoptotic index in any cell group and serum progesterone concentrations.     

Investigation of individual and group variability in estrous cycle characteristics in female Asian elephants (Elephas maximus) at the Oregon Zoo

Evaluating ovarian cycle activity through longitudinal progestagen monitoring is important for optimizing breeding management of captive elephants and understanding impact of life events (births, deaths, and transfers) on reproductive function. This study summarized serum progestagen profiles for eight Asian mainland elephants (Elephas maximus indicus) and one Bornean elephant (E. maximus borneensis) at the Oregon Zoo over a 20-yr interval, and represents the longest longitudinal dataset evaluated to date. Estrous cycle characteristics were more varied than previously reported for this species, with an overall duration of 12 to 19 wk, luteal phase duration of 4 to 15 wk, and follicular phase duration of 2 to 12 wk. In general, there was more cycle variability across than within individual elephants. Compared with other elephants in the group, the Borneo female exhibited consistently longer cycle lengths, higher progestagen concentrations, and greater cycle variability; however, it is not known if this represents a subspecies or an individual difference. Cycle durations did not appear to change over time or with age, and the first pubertal cycle was similar to subsequent cycles. Variability in duration of the follicular phase was greater than that of the luteal phase. In addition, there was a significant negative relationship between luteal and follicular phase durations, suggesting a possible regulatory role of the follicular phase in maintaining a relatively consistent cycle duration within individuals. Overall, we found these elephants to be highly resilient in that major life events (births, deaths, and changes in herd structure) had minimal effect on cycle dynamics over time. In conclusion, the higher range in cycle phase characteristics is likely because of the larger number of elephants studied and longer duration of longitudinal monitoring, and may be more representative of the captive population as a whole. Furthermore, identification of significant interanimal variability suggests that understanding the complexities of herd reproductive characteristics could facilitate development of more effective institution-specific breeding management strategies.     

Factors affecting gestation length and estrus cycle characteristics in Spanish donkey breeds reared in southern Spain

This paper investigated gestation length and estrus cycle characteristics in three different Spanish donkey breeds (Andalusian, Zamorano-Leones, and Catalonian) kept on farm conditions in southern Spain, using data for ten consecutive breeding seasons. Gestation length was measured in 58 pregnancies. Ovarian ultrasonography was used to detect the ovulation, in order to ascertain true gestation length (ovulation-parturition). Pregnancy was diagnosed approximately 14-18 d after ovulation and confirmed on approximately day 60. Average gestation length was 362 ±15.3 (SD) d, and no significant differences were observed between the three different breeds. Breeding season had a significant effect (P < 0.01), with longer gestation lengths when jennies were covered during the early period. Breed, age of jenny, year of birth, foal gender, month of breeding, and type of gestation had no significant effect on gestation length.After parturition, foal-heat was detected in 53.8% of the postpartum cycles studied (n = 78), and ovulation occurred on day 13.2 ± 2.7. The duration of foal-heat was 4.7 ±1.7 d, with a pregnancy rate of 40.5%.When subsequent estrus cycles were analyzed, the interovulatory interval (n = 68) and estrus duration (n = 258) were extended to a mean 23.8 ± 3.5 and 5.7 ± 2.2 d, respectively. Both variables were influenced by the year of study (P < 0.03 and P < 0.001), whereas month and season of ovulation (P < 0.005 and P < 0.009, respectively) affected only interovulatory intervals. Estrus duration was significantly longer than that observed at the foal-heat (P < 0.006), and the pregnancy rate was 65.8%.This study provides reference values for true gestation length and estrus cycle characteristics in Spanish jennies. Breeding season affected gestation length in farm conditions. Also, seasonal influence was observed on the length of the estrus cycle (i.e., interovulatory interval), although foal-heat was not affected by environmental factors.     

Sexual behavior and ejaculate characteristics in Pêga donkeys (Equus asinus) mounting estrous horse mares (Equus caballus)

The objectives were to (i) characterize sexual behavior of donkey stallions (jacks; Equus asinus) during on-farm semen collection using estrous horse mares (mares; Equus caballus); (ii) compare behavior of young (less experienced) versus older (more experienced) jacks; (iii) determine whether semen suitable for artificial insemination (AI) could be collected using mares; and (iv) determine the suitability of using mares in field collection of semen from jacks. Six Pêga jacks (3.5 to 16 yr old), previously conditioned to breed mares, were used. Mount mares were confirmed in estrus by a teaser horse stallion (stallion) and a jack. Semen was collected with an artificial vagina, at intervals of 48 to 72 h (180 collections). The mean ± SD (young [3.5 yr] vs. old [14 to 16 yr]) were Flehmen response frequency, 7.4 ± 5.8 (8.1 ± 3.0 vs. 7.0 ± 2.0); number of mounts without erection, 1.1 ± 1.3 (2.1 ± 1.4 vs. 1.2 ± 0.4, P < 0.05); latency from first exposure to mare to full erection on the ejaculatory mount, 18.3 ± 17.7 min (25.3 ± 21.3 vs. 12.2 ± 6.2, P < 0.05); latency from erection to insertion, 5.1 ± 3.5 sec (5.3 ± 3.8 vs. 4.8 ± 3.2); and duration of copulation from insertion to dismount after ejaculation, 25.4 ± 7.8 sec (22.1 ± 2.9 vs. 28.1 ± 9.3). In all jacks, sexual behavior was generally normal, with the notable absence of open mouth behavior. Mare estrous behavior was markedly less intense than that in the presence of a stallion and usually absent. Semen characteristics were gel free volume, 47.3 ± 28.7 mL; gel volume, 71.8 ± 54.8 mL; total motility, 84.3 ± 6.0%; progressive motility, 74.3 ± 74.5%; sperm vigor, 3.9 ± 0.5 (scale 1 to 5); sperm concentration, 253 × 10⁶ cells/mL; and total number of sperm, 10.3 × 10⁹ cells. Copulation duration was significantly correlated with gel free volume (r = 0.9) and gel volume (r = 0.7). We concluded that (i) the sexual behavior of jacks during semen collection using mares was similar to that reported for natural mating to jennies, (ii) precopulatory and copulatory behavior for the young (less experienced) jacks and older (more experienced) jacks were generally similar (except number of mounts without erection and latency to full erection); (iii) semen obtained using mares as stimulus and mount females was similar to that reported with estrous jennies; and (iv) semen collection from previously conditioned jacks, using estrous mares, was appropriate for field collection of semen.     

Antioxidant capacity of follicular fluid in relation to follicular size and stage of estrous cycle in buffaloes

The present study was designed to evaluate the changes in the concentrations of different antioxidants, such as glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), and catalase (CAT), in the follicular fluid collected from different follicular size categories in relation to stage of estrous cycle in buffaloes. In addition, malondialdehyde (MDA) as an indicator for lipid peroxidation was also estimated. Fifty pairs of buffalo ovaries were collected from a local slaughterhouse. Based on ovarian structures, the cycle was divided into follicular and luteal phase. The follicles on each pair were classified into three groups; small (≤3 mm), medium (4-9 mm) and large (≥10 mm). The concentrations of SOD, CAT, GSH, and GR in the follicular fluid of each group as well as MDA were estimated. Results indicated that there was a significant decrease (P < 0.05) in the average numbers of small follicles obtained at the follicular phase than those obtained at the luteal phase of the cycle. However, the mean numbers of the large sized follicles was significantly increased (P < 0.05) in the follicular phase than in the luteal phase. Large follicles obtained at the luteal phase had a significantly higher (P < 0.05) concentration of GSH than that obtained from small ones. A significant (P < 0.05) effect of follicular size on GR concentrations was observed. The concentration of SOD tended to be higher in large follicles obtained at the follicular phase than that collected at the luteal phase (56.7 ± 3.7 vs. 28.1 ± 6.7 U/mL, respectively). On the contrary, a significantly higher concentration (P < 0.05) of SOD was recorded in small follicles as compared with medium and large follicles collected at the luteal phase. CAT concentrations did not significantly differ among different follicular sizes between follicular and luteal phases as well as within each phase. Malondialdehyde concentration was significantly decreased (P < 0.05) in the follicular fluid obtained from small follicles collected at the follicular phase compared with those obtained at the luteal phase. In conclusion, the present study showed that the concentrations of enzymatic antioxidants except for CAT vary according to the follicle size and the stage of the estrous cycle suggesting their possible role in the process of follicular development during estrous cycle in buffaloes.     

A stereological study of medium antral follicles during the bovine estrous cycle

Stereological methods were applied to bovine cumulus-oocyte complexes (COCs) in order to characterize them quantitatively during the estrous cycle. COCs from medium (4-8mm) antral follicles with a compact and complete cumulus mass, and with an uniform or a non-visible ooplasm were aspirated from ovaries of Holstein-Friesian cows, fixed in glutaraldehyde, randomly embedded in glycol-methacrylate, and sectioned at 20 microm. The unbiased nucleator principle was used for estimating the mean volumes of complexes, oocytes, cumulus cells, and nuclei of oocytes and cumulus cells. The thickness of the zona pellucida and the relative numerical percentages of the several morphological types (C1-C3) of cumulus cells were also evaluated. The optical disector procedure was used for cumulus cell sampling. Quantitative data show that COCs appear heterogeneous for all studied parameters. From metestrus to proestrus the volumes of COCs and oocytes remained constant, the volumes of oocytes and oocyte nuclei were correlated, the thickness of the outer zona pellucida decreased, and the relative numerical frequency of follicular type C3 cells increased. Results suggest that COCs from distinct estrus stages are structurally different, with type C3 follicular cells gradually differentiating from cell types C1 and C2.     

Studies on the relationship between paracoccidioidomycosis in ddY mice and their estrous cycle

The relationship between paracoccidioidomycosis in ddY mouse and its estrous cycle was studied. Adult ddY mice of both sexes were used as experimental animals. Estrous cycle of female mice was examined before inoculation of Paracoccidioides brasiliensis yeast cells and mice were divided into 5 groups such as proestrus, estrus, metestrus-I, metestrus-II and diestrus. Each mouse was inoculated intravenously with 10⁶ P. brasiliensis yeast cell units and sacrificed on day 28 after inoculation. Their internal organs were cultured, and in addition, their histopathologies were studied. As a result, there was no difference in the organ cultures among the male and the female mice of 5 groups. However, histopathologically, the female groups at estrus, metestrus-I and metestrus-II were affected more severely than the male group, and the susceptibility of the female mice to the fungus was closely related to their estrous cycles.Abbreviations BHI-D brain heart infusion agar supplemented with 1.0% of anhydrous dextrose - PAS periodic acid-Schiff techniques - PBS phosphate buffered saline solution - SD standard deviation     

Diffusion barriers in the vaginal epithelium during the estrous cycle in guinea pigs

Summary In the present study the permeability barriers of the multilayered vaginal epithelium were examined using tracer perfusion techniques, freeze-fracture and thin sectioning. During diestrus and proestrus the upper layers of mucified epithelial cells exhibit tight-junctional belts, which restrict tracer molecules such as lanthanum and horseradish peroxidase. When the highly mucified cells begin to degenerate toward the end of proestrus the underlying epithelium is already keratinized as typical for estrus. The keratinized epithelial cells have a tight-junctional network that joins the basal plasma membranes with the apical membranes of subjacent cells and blocks paracellular diffusion of the tracer molecules. During conversion of the cornified epithelium to a mucified epithelium in metestrus the intercellular space of the epithelium is stained by tracer molecules even though tight-junctional belts can be observed.These results indicate that during cyclic changes of the vaginal epithelium tight junctions can, in general, be considered for the restriction of paracellular diffusion. In metestrus, however, junctions become functionally leaky although they remain morphologically intact.Intercellular lipids, which are normally common in cornified epithelia, are extremely rare and cannot constitute an effective barrier to diffusion in the vagina of the guinea pig. The significance of a strategy that bases the regulation of the permeability on tight junctions rather than on intercellular lipids is discussed.     

Effect of season on estrous cycle of Yankasa sheep

An investigation was conducted to establish the effects of harmattan and hot-dry season on estrous cycle length, onset, and duration of estrus in Yankasa sheep indigenous to the Nigerian guinea savanna zone. Mean cycle lengths were 16.8 +/- 0.58 and 16.4 +/- 0.53 days during harmattan and hot-dry seasons, respectively; short cycles, 5-13 days, and long cycles, 21 to 30 days, were observed during both seasons. During the harmattan season, 57.1% of estrus began at night while 70% started at night during the hot-dry season. The duration of normal estrus observed during the harmattan, 33.6 +/- 5.87h, significantly decreased (P0.05) during the hot-dry season (24.0 +/- 5.45h). It is suggested that twice daily observation at 12-hour intervals will suffice to detect estrus in this breed of sheep.     

A collaboration of aquaporins handles water transport in relation to the estrous cycle in the bitch uterus

Fluid movement through uterine cell membranes is crucial, as it can modulate the tissue imbibition pattern in the different phases of the estrous cycle. To gain insight into the mechanisms underlying steroid-controlled water handling, the presence and distribution of aquaporins (AQPs), integral membrane channel proteins permitting rapid passive water movement, was explored in bitch uterine tissues. Immunohistochemistry and Western immunoblot analysis were used to study the presence of AQP1, AQP2, and AQP5 in the layers of the bitch uterine wall during the different estrous phases. Presence of endothelial nitric oxide-generating enzyme NO synthase (NOS3) was also investigated, as it is known that the vasodilator NOS3 might be involved in the development of uterine edema. The results demonstrated the following: (1) AQP1, AQP2, and AQP5 were present in the uterus of cycling bitches. (2) AQP1 was localized within uterine mesometrial, myometrial, and endometrial blood vessels and in the circular and longitudinal layers of myometrium. AQP1 localization and expression were unaffected by the estrous cycle. (3) The estrogenic milieu was probably at the basis of AQP2 expression in the glandular and luminal epithelium of the endometrium. (4) AQP5 water channels were present in the apical plasma membrane of uterine epithelial cells in coincidence with plasma progesterone increase. (5) NOS3 was localized in the myometrial and epithelial tissues as well as in blood vessels indicating a contribution of this vasoactive peptide to the uterine imbibition processes. Thus, we can hypothesize that a functional and distinctive collaboration exists among diverse AQPs in water handling during the different functional uterine phases.     

Lymphocyte number and distribution in the rat uterine epithelium during estrous cycle and early pregnancy

Summary The number of intraepithelial lymphocytes (IEL) in the luminal and glandular epithelium of the uterus of virgin rats was analysed in diestrus, proestrus and estrus, and in nulliparous rats on days 5, 7 and 9 of pregnancy. IEL number was calculated either with respect to the number of epithelial cells or to the length of epithelium section. It was found that in diestrus, the number of IEL was, on average, 3.7 per 100 luminal epithelial cells or 6.7 per 1 mm of epithelium section, whereas in proestrus, it decreased to 0.9 and 1.2 IEL, respectively. On day 5 of pregnancy (before implantation) the number of IEL decreased further to 0.45 per 100 luminal epithelial cells or 0.9 per 1 mm of epithelium. On days 7 and 9 of pregnancy, IEL number further decreased in implantation sites, whereas in interimplantation sites it remained at the level calculated for day 5 of pregnancy. The population of uterine IEL consisted of small (82–99%) and large (1–18%) lymphocytes. In all stages of the estrous cycle, IEL occurred with a frequency of 68–87% in the basal region, 8–20% in the middle region and 4–12% in the apical region of the luminal epithelium width.     

The effect of the estrous cycle and estrogen on the release of immunoreactive alpha-melanocyte-stimulating hormone

Circulating levels and tissue content of alpha-MSH were measured on the morning of various days of the estrous cycle, and on the afternoon of proestrus in freely moving conscious rats. No surges of alpha-MSH were detected by RIA in the morning of various days of the cycle. The neurointermediate lobe content of alpha-MSH was slightly elevated on diestrus 1 as compared to the levels on diestrus 11 and proestrus but not to estrous levels. No changes in alpha-MSH content were detected in the anterior pituitary, the median eminence, mediobasal hypothalamus and the preoptic area at various stages of the estrous cycle. Plasma alpha-MSH levels were slightly elevated at 1500 hr of proestrus which was followed three hours later by a decline. This profile of plasma alpha-MSH on the afternoon of proestrus was reproduced by the SC administration of estradiol benzoate to long-term ovariectomized rats. These data suggest that, contrary to the results obtained by bioassay of alpha-MSH no surges of alpha-MSH occur on any day of the cycle, although a slight elevation on the afternoon of proestrus was detected. The altered pattern of release of this peptide on the afternoon of proestrus may be induced by estrogen.     

Molecular mechanism of cell proliferation in rodent uterus during the estrous cycle

The rodent uterus is a widely studied target tissue for sexual steroid hormone action. The aim of the present study was to assess the molecular mechanism that participates in the initiation of cell proliferation of the rat uterine epithelial cells during the estrus (E)–metestrus (M) transition. Cell proliferation, ERα, c-fos, cyclin D1 and D3, cdk4, and cdk6 proteins were assessed in these animals by immunohistochemistry. Estradiol (E₂) and progesterone (P₄) plasma levels were assessed by RIA. The results indicate that the glandular epithelium starts to proliferate at 21:00 h on estrus day, and initiates at least 3 h before the luminal epithelium does. Fos expression was markedly increased during the afternoon of estrus day, and its increase was in parallel to ERα expression. Interestingly, both, cyclin D1 and D3 were abundantly expressed in the luminal and glandular epithelia, and nuclear immunolabelling of cyclin D1 and D3 precedes BrdU incorporation in the cell. cdk4 and cdk6 were localized in the nuclei in both epithelia throughout the studied time course. In addition, cdk4 was more abundant throughout estrus and metestrus days than cdk6. The overall results indicate that ERα, Fos and cyclins D1 and D3, cdk4 and cdk6 are expressed in both glandular and luminal epithelia of the rat uterus during the E–M transition. In conclusion, there is a good correlation between sequential expression of these proteins and cell cycle progression in the rat uterine epithelial cells during the estrous cycle. However, the differences observed in the cellular localization, time course of expression and the cellular types that express both cyclins between physiological and pharmacological conditions, demonstrated different mechanisms of regulation and should be due to the complex hormonal milieu during the estrous cycle.     

Plasma follicle stimulating hormone and luteinizing hormone patterns during the estrous cycle of ewes

Fifteen Suffolk ewes were used in three experiments to compare plasma follicle stimulating hormone (FSH) and luteinizing hormone (LH) patterns during the estrous cycle and to determine whether FSH levels undergo changes in pulse frequency. Luteinizing hormone changed inversely with progesterone levels whereas FSH and progesterone concentrations revealed no obvious relationship. Unlike LH, FSH levels did not pulsate during the follicular phase. Higher FSH levels were detected on days 1, 6 and 12 and lower levels on days 0, 4 and 16. Coincident preovulatory LH and FSH surges were observed and this was the only time FSH and LH levels appeared to be jointly controlled.