Morphologic changes due to human chorionic gonadotropin in the rat testis: Role of vascular endothelial growth factor

Introduction: Human chorionic gonadotropin (hCG) has an important role in the regulation of reproductive system and a high capacity to stimulate angiogenesis by secretion of vascular endothelial growth factor (VEGF). It could be also used for medical treatment of cryptorchidism, as it stimulates Leydig cells to produce testosterone, but its use has been discussed. Aim of the study was to evaluate the testicular morphological changes due to cyclic administration of hCG possibly related to VEGF effect. Methods: 80 pre-puberal male Sprague-Dawley rats were randomly divided in two groups hCG (50 U/I/ Kg) (n = 55/S) and placebo (n = 25/C) treated. The animals were sacrificed respectively after one (S1 n = 10;C1 n = 5), two (S2 n = 10;C2 n = 5) and three weeks of hCG or placebo treatment (S3 n = 35;C3 n = 15);a blood serum was obtained for evaluation of plasmatic testosterone. The testis, prostates, epididymides and seminal vesicles were drawn and weighted. Specimens were fixed in formalin and coloured by hematoxylin-eosin. Morphometric analysis of Leydig cell number, nuclear area and perimeter was carried out. Immunoistochemistry for VEGF has been performed with the avidin-biotin peroxidase technique. Statistical differences between groups, each of different age, were calculating using Student’s T-test. Results: After the first week of hCG treatment it has been observed an increased testicular weight and volume (+/﹣6%) in experimental respect to control group. This result was not observed after the second and third week of treatment. Significant variations were also observed in the mean prostate weight, whereas mean seminal vesicles weight was significant in the S2 and S3 groups respect to control. In the hCG treated rats it has been observed a poor differentiation of the seminiferous epithelium, with high Leydig cell evidence and increased intertubular eosinophilic material. An interstitial edema was demonstrated without inflammatory cell changes. Morphometric analysis confirmed an increased number of Leydig cells that were always strongly immunostained for VEGF. Low-medium reactivity was present in spermatogonial, peritubular, endothelial and Sertoli cells. Testosterone plasmatic values showed a statistically significant and progressive increase. Conclusions: hCG treatment clutters the germinative line and modifies VEGF testicular expression, suggesting its role in male reproductive system. The increased number of Leydig cells and the immunostaining for VEGF suggests an intense interstitial and metabolic arrangement. The morphologic modifications induced by hCG treatment and its influence on testis function raise the question about its use in the medical treatment of cryptorchidism.

Conjunctival cytological examination,bacteriological culture,and antimicrobial resistance profiles of healthy Mediterranean buffaloes(Bubalus bubalis) from Southern Italy

Objective: To assess normal conjunctival cytological and bacteriological/fungal flora features in the Mediterranean buffalo(Bubalus bubalis).Methods: Swabs were taken from the inferior conjunctival sac of both eyes of 57 healthy female buffaloes aged 24–36 months, with no evidence of ocular disease, farmed in Campania region(Southern Italy), for microbiological analysis. Conjunctival eye specimens of both eyes were subsequently obtained by a cyto-brush, for cytological analysis.The antimicrobial susceptibility of bacterial isolates was also determined using the diskdiffusion method on Mueller Hinton agar plates.Results: Cytological examination of conjunctival swab specimens(114 eyes) revealed epithelial cells(basal, intermediate, columnar and super ficial) in all samples, whereas neutrophils, lymphocytes and plasma cells were present in 70%, 10% and 2% of samples,respectively. Microorganisms, for a total of 261 aerobic bacteria and 6 fungi, were isolated from 112/114 conjunctival samples [98.25%; 95% con fidence interval(CI): 93.18–99.70]. Only two conjunctival swabs did not yield bacteria and/or fungi(2/114, 1.75%;95% CI: 0.30–6.82). Gram-positive aerobes were most commonly cultured(181/261,69.35%; 95% CI: 63.31–74.81), with Enterococcus faecium and Staphylococcus lentus predominating. Escherichia coli was the most frequently isolated as Gram-negative bacteria(80/261, 30.65%; 95% CI: 25.19–36.69). The antimicrobial resistance patterns of the isolated bacteria showed amoxycillin/clavulanic acid and cephalothin as the least sensitive antibiotics for both Gram-positive and Gram-negative bacteria.Conclusions: These results provided first information on normal conjunctival ocular micro flora and cytological features in Mediterranean buffalo.