Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

Effects of monoamines formed in the cecum of horses on equine digital blood vessels and platelets

OBJECTIVE: To determine in vitro vasoactive potency of monoamines formed in the cecum and found in the systemic circulation of horses. SAMPLE POPULATION: Segments of digital blood vessels obtained from 6 healthy mixed-breed horses and ponies euthanatized at an abattoir and platelets isolated from 4 healthy ponies. PROCEDURE: Paired rings of digital artery and vein from the same horse were examined, and isometric tension was recorded. Concentration-response curves for tryptamine (TRP), tyramine (TYR), phenylethylamine (PEA), isoamylamine (IAA), and isobutylamine (IBA) were obtained. Vasoconstrictor mechanisms were investigated for TRP and TYR by the use of antagonists. Washed platelets loaded with [3H]-5-hydroxytryptamine (5-HT) were incubated with monoamines; the amount of radioactivity displaced after 30 minutes was estimated. RESULTS: TRP, TYR, and PEA were potent constrictors of arteries and veins, with TRP and TYR being more potent in veins than arteries. Constrictions induced by TYR were inhibited by benextramine (alpha-antagonist) and nisoxetine (neuronal-uptake blocker), whereas TRP responses were inhibited by ketanserin (5-HT receptor antagonist). All 5 amines displaced 5-HT from platelets with the order of potency being TYR > TRP > PEA > IAA > IBA. CONCLUSIONS AND CLINICAL RELEVANCE: Amines from the equine cecum cause digital vasoconstriction. The most potent (TRP and TYR) cause selective venoconstriction. Tyrosine activates predominantly alpha-adrenoceptors through the release of neuronal norepinephrine, whereas TRP activates 5-HT receptors. All amines tested released 5-HT from platelets. Amines formed in the cecum and released into the systemic circulation warrant additional investigation as trigger factors for digital ischemia and subsequent laminitis.     

The function, composition and analysis of cerebrospinal fluid in companion animals: part II - analysis

Accurate analysis of cerebrospinal fluid (CSF) provides a wide range of information about the neurological health of the patient. CSF can be withdrawn from either of two cisterns in dogs and cats using relatively safe techniques. Once CSF has been collected it must be analysed immediately and methodically. Evaluation should consist of macroscopic, quantitative and microscopic analyses. As part of a quantitative analysis, cell counts and infectious disease testing are the most important and potentially sensitive indicators of disease. Although certain pathologies can be described, microscopic analysis will rarely be specific for any disease, emphasising the adjunctive nature of this diagnostic modality.     

A Bayesian meta-analysis of the effects of administering an intra-vaginal (CIDR) device in combination with other hormones on the reproductive performance of cycling,anoestrous and inseminated cows

AIMS: To evaluate the effectiveness of treatment programmes that included controlled internal drug-releasing (CIDR) devices containing progesterone (P4) in improving synchrony of oe- strus, and conception and pregnancy rates in cycling, anoestrous and inseminated dairy cows, using meta-analysis. To describe the difference in response between cycling and anoestrous cows to CIDR-based synchrony programmes.

METHODS: Scientific papers written in the English language between 1989 and 2002 that investigated the effects of treat- ment programmes including CIDR devices on reproductive per- formance in dairy heifers or lactating dairy cows were identified using a computerised literature search. The criteria for inclusion incorporated evidence that treatment allocation was completely randomised; the population studied was lactating dairy cows; and that data were available on submission, conception and pregnancy rates and their associated measures of variability. Re- productive outcomes from 25 synchrony trials (total n= 11,058 cows) were analysed. Summary measures of the effect of treat- ment on reproductive outcome were assessed using fixed- and random-effects Bayesian meta-analysis models.

RESULTS: Treatment programmes including a CIDR device increased the risk of submission in cycling cows (predicted Bayesian RR=2.86, 95% credible interval 1.46–5.67). Compared with controls, synchrony programmes including CIDR devices in cycling dairy cows had no effect on the risk of conception to first service post-treatment (predicted Bayesian RR=1.00, 95% credible interval=0.80-1.24). Compared with controls, synchrony programmes including CIDR devices had no effect on the risk of pregnancy throughout the mating period (pre- dicted Bayesian RR=1.02, 95% credible interval=0.89-1.17). In anoestrous cows, CIDR treatment had no effect on the risk of conception to first service post-treatment and no effect on the risk of pregnancy throughout the mating period, compared with anoestrous, untreated controls (predicted Bayesian RR=0.91 and 0.97, respectively; 95% credible interval=0.68–1.26 and 0.59-1.60, respectively).

CONCLUSION: The results of this meta-analysis showed that synchrony programmes using CIDR devices combined with other hormones reliably enhanced submission rates in lactat- ing dairy cows. The relatively small number of trials with data suitable for analysis and the heterogeneity of results at the indi- vidual trial level limited our ability to confirm either a beneficial or deleterious effect of treatment on conception or pregnancy rates. Further randomised, controlled trials to evaluate the ef- fectiveness of this form of reproductive therapy in commercial dairy farms are needed.     

Acrosomal and viability status of bovine spermatozoa evaluated by two staining methods

Artificial insemination with frozen-thawed spermatozoa is commonly used in cattle breeding. A simple and fast procedure is needed for routine evaluation of the acrosomal status of frozen-thawed bovine sperm. Therefore, the purpose of this study was to test two staining procedures used to determine the viability and integrity of acrosome of frozen-thawed bovine spermatozoa. Double staining and Hoechst/FITC-Pisum sativum agglutinin (FITC-PSA) labelling were tested for evaluating the viability and acrosome reaction induced by calcium ionophore of bull spermatozoa. In our experiments no significant differences were detected in the frequency of acrosome-reacted sperm either by double staining (37.98%) or by FITC-PSA labelling (39.33%). The viability of sperm stained by the double staining method was 67.17%, and a higher portion of viable sperm (82.67%) was observed by staining with the Hoechst procedure (P < 0.01). On the basis of the results obtained it is concluded that both methods can be used for detecting the acrosome reaction of frozen-thawed bovine spermatozoa.     

Effect of gallium on photosynthetic pigments and peroxidase activity of Chlorella pyrenoidosa

Green algae (Chorella pyrenoidosa) were grown in iron‐deficient and normal iron supplied nutrient solution and treated with 1 uM gallium. Treatment with gallium enhanced the growth rate and chlorophyll content of algae cultures. Significant changes were recorded in individual photosynthetic pigment composition of treated cultures. In both iron‐deficient and normal iron supplied cultures, gallium treatment enhanced peroxidase enzyme activity. Partially purified peroxidase showed different kinetic properties in the samples treated with gallium. An iron‐gallium interaction is proposed that affects chlorophyll metabolism and peroxidase activity in green algae.