Effects of intratesticular vs intraepididymal calcium chloride sterilant on testicular morphology and fertility in dogs.

Abstract Background Both stray and free-roaming owned dogs contribute to the serious global dog overpopulation problem. Many dog owners are unwilling to have their pet castrated for various reasons, including a reluctance to have their dog's behavior changed. A non-surgical method of sterilizing both stray and owned dogs would help to prevent unwanted litters. Previous studies have shown that intratesticular injection of calcium chloride dihydrate (CaCl2) in alcohol is a promising and cost-effective alternative to surgery for stray dogs, with testosterone significantly decreased and sexual activity eliminated. The aim of this study was to compare the use of a solution of 20% CaCl2 in 95% ethanol injected into the testicles or into the head of the epididymis. Methods A total of 148 dogs divided into 4 groups (2 experimental and 2 control) were respectively injected with CaCl2 or saline solution into the testicle or epididymal head (ultrasound-guided). The animals were examined at 0, 3, 6, and 9 months for sperm quality, concentration of testosterone in serum, and side effects; at 0 and 5 months with contrast-enhanced ultrasound (CEUS) to enhance the morphological aspects/alteration of the testicular parenchyma or epididymis; and at 9 months when all were castrated for histological examination. Results All dogs treated with CaCl2 became sterile with azoospermia achieved over the 9-month study. The concentration of testosterone in serum significantly decreased following intratesticular treatment with CaCl2. No adverse effects were noted. Conclusions A single, bilateral intratesticular injection of 20% CaCl2 in 95% ethanol was confirmed to be a reliable method for induction of sterilization in male dogs. The approach showed long-term efficacy and may reduce sexual behavior, with the additional benefits of low-cost and ease of use, making this nonsurgical method appropriate for use in stray dogs. Sterility was also achieved if injected in the head of the epididymis but no significant decrease in serum concentration of testosterone occurred. Moreover, performing the intraepididymal injection into the epididymal head was as time consuming as orchiectomy. This approach may be optimal for use in owned dogs where anatomical integrity and testosterone maintenance is preferred by the owner

Therapeutic Ultrasound as a Potential Male Dog Contraceptive: Determination of the Most Effective Application Protocol

Contents: Ultrasound is one of the most promising forms of non-invasive contraception and has been studied in several animal models. The objective of the current investigation was to determine the most practical and effective application protocol for dog sterilization. A total of 100 dogs were divided into five equal groups. Group A received 5-min applications three times performed at 48-hr intervals and covering the entire testicular area at frequency of 1 MHz; Group B received 5-min applications three times performed at 48-hr intervals over the dorso-cranial area of the testis at frequency of 3 MHz; Group C received three sequential 5-min applications (at 5-min intervals between applications) covering the entire testicular area at frequency of 1 MHz; Group D received 15-min applications two times performed at 48-hr intervals and covering the entire testicular area at frequency of 1 MHz. The experimental groups' ultrasound had an intensity of 1.5W/cm2. The Control Group had the same procedure as Group A, but with the transducer switched-off. Dogs were surgically castrated 40 days following the treatment for histological examination. Azoospermia, testicular volume reduction and apparently irreversible testicular damage were achieved by Group A. No effects were noticed in the other groups. Testosterone levels remained within physiological range with all application protocols. A regimen of three applications of ultrasound at 1 MHz, and 1.5 W/cm2, lasting 5 min with an interval of 48 h was effective as permanent sterilization in the dog without hormonal impact

Pesticides: An Overview of the Current Health Problems of Their Use

The necessity to produce increasing quantities of food to meet the demand of ever-increasing populations has led to the massive use of pesticides in agriculture. Their massive consumption derives from the need to subtract food from competition of fungi, bacteria, and insects and agricultural land from the in- vasion of competing weeds. Modern agriculture floats on an ocean of syn- thetic chemical compounds used to crop protection and food preservation during storage. Presently, worldwide about 2000 active ingredients have been listed, categorized into about 60 classes of chemicals. In the world, the overall pesticides, used in agriculture, are about 4.1 million tonnes/years. The exposure of animals and humans to pesticides could be related to neurotoxicity, endocrine disruption, liver and kidney damage, cancer, reproductive effects (sperm abnor- malities, decreased fertility, fetal growth retardation, birth defects, or sponta- neous abortion), and growth modifications. However, the knowledge about their potential damage to the organs of reproduction, breastfeeding, and in- fants is still largely insufficient. It is necessary to improve new methods to identify and regulate the long-term safety of pesticides use, to protect the en- vironment and health, and to eliminate the adverse effect of pesticides on the environment and non-target organisms. The assessment of possible risks of old and new pesticides must be increased to include the possible effects on reproduction on animals and pregnancy loss even after a long time. In the meantime, it is necessary to minimize the use of synthetic chemical compounds by increasing the use of biological systems

Clomazone: one of the latest really low toxic multi-crop herbicides?

To reduce pollution and risks of pesticides, numerous studies and research are carried out to identify new degradable compounds, identify more effective and less dangerous formulations. For this reason, hundreds of new compounds and formulations are continuously available every year. Use, programs and controls of these chemical compounds are not uniform all over the world. In some poor or developing countries, means to control exposures and risks, associated with these substances, are not available or limited. The main active components of herbicides, registered in the world in 2019, were clomazone and isoproturon, used as single agents or mixed with other components. The use of Clomazone, in particular, has grown in many countries, but in China and other emerging economies in the recent past there has not been the expected expansion, despite the growing world demand. In fact, many Chinese producers have ceased the production of this compound for its higher production costs than other substances. However, analysts expect an increase, for the next years, of clomazone consumption for various reasons, first of all, because it is considered much less toxic than other products. However, in the absence of indications deriving from long periods use, it seems evident the need to avoid recourse to the massive use of this herbicide by carefully observing the indications and precautions provided by the appropriate international bodies (US EPA, EFSA, etc.). As regards the studies on the effects on the reproductive functions of both terrestrial and marine animal organisms, it is evident the need to increase their number, until now completely insufficient

Unusual giant cyst as a consequence of the ovarian remnant syndrome in a bitch.

An 9 years old German shepherd bitch was presented to the Veterinary Hospital of the University of Bari "Aldo Moro" with a 1 year history of enlarged abdomen. The bitch was neutered 3 years before and no signs of heat were seen since then. The owner claimed a progressive increase in abdominal volume that had started about 1 year earlier and progressed faster in the last 2 months when there also was a deterioration of the general conditions with dysorexia. It worsened in anorexia for about 2 days. On physical examination the bitch was thin, depressed and reluctance to move with a distended and swollen abdomen. At superficial palpation the abdominal wall resistance was detected, the rigidity was due to an intra-abdominal fluid collection. A complete blood and urine tests showed nothing relevant. The ultrasonographic examination showed the presence of a spider-web-like structure with septa and an anechoic content that occupied most of the abdominal cavity displacing the viscera. We proceeded with an exploratory laparotomy. The opened abdomen revealed the presence of a cystic formation that occupied most of the abdominal cavity (about 25 cm in diameter) developed on a remnant of the right ovary and containing about 13 lt of serum-hematic fluid and many adhesions between abdominal organs and tissues. The presence of other cysts on a further left ovarian residue was also highlighted: one with a diameter of about 10 cm and others smaller than about 5 cm. Fragments of right ovarian cysts and uterus were stored in 2% buffered formalin to be subjected to histological examination. The subject recovered from anesthesia was discharged with the following therapy: amoxicillin clavulanate (12.58 mg/kg bid for 10 days) and robenacoxib (1 mg/kg sid for 5 days). The histological examination of the cysts revealed that the wall had the internal theca replaced by fibrous tissue, several layers of degenerated granulosa cells. This finding suggest a follicular stromal cyst. Uterine tissue showed signs of cystic endometrial hyperplasia. At 10 days post-surgery check-up the bitch appeared to be in good health with no signs of depression and pain. She was completely recovering and gaining weight. The owner claimed that the bitch had started feeding again the day post surgery and had been improving day by day. Usually the subjects with ovarian remnant syndrome (ORS) manifest the typical signs of estrus, such as vulvar serohematic discharge, receptivity to the male, nymphomania, and kyphosis due to abdominal pain. Additional clinical signs are: endometrial cystic hyperplasia, pyometra and infrequently diabetes mellitus (1). The peculiarity of this clinical case is represented by the giant dimension of the follicular cyst and the presence of non-specific clinical signs due to the compre

The Effect of Dietary Supplementation of Vitamin E, Selenium, Zinc, Folic Acid, and N-3 Polyunsaturated Fatty Acids on Sperm Motility and Membrane Properties in Dogs

Sub-fertility represents a common challenge in canine reproduction. Different protocols, supplementing daily given quantities of micronutrients, were investigated to improve poor sperm concentration and/or function, which represent breeding major constraining factors in the canine species. Little information is available for dogs concerning the effect of a daily supplementation with a complex of vitamin E, zinc, selenium, folic acid, and n-3 polyunsaturated fatty acids (PUFA) on semen quality. Thus, the present study investigated this effect on semen motility and sperm membrane properties. Serial semen analyses from fourteen healthy normospermic dogs, fed with the same commercial diet, were performed on Days 0 (T0), 30 (T30), 60 (T60), and 90 (T90). Seven dogs were randomly included in the treatment (T) group, receiving a supplementation of vitamin E, zinc, selenium, folic acid, n-3 PUFA; and seven other subjects composed the control (C) group. Total Sperm Count (TSC), Computer-Assisted Sperm Analysis (CASA) indexes, mortality, and functional membrane integrity were assessed. The ANOVA compared results between groups and sampling times (p < 0.05). From T60, the supplementation significantly improved TSC, progressive motility, functional membrane integrity, and decreased mortality. Present results lead us to consider ameliorative effects of a two-month healthy diet supplementation on canine spermatozoa. The positive effects of the described balanced integration of micronutrients on sperm motility and prevention of oxidative stress should be considered, especially when decreased seminal parameters may result from inadequate intake, reduced absorption, increased losses or demand, or to attenuate the impact of age

Ejaculation Effect on Canine Prostatic Specific Esterase Serum Concentration

Canine prostatic diseases are usually asymptomatic in their onset and often identified in advanced stages. Canine prostatic specific esterase (CPSE) represents an early serum marker for prostatic diseases, also in asymptomatic dogs. The present study aimed to identify the effects of ejaculation on serum CPSE. Twenty adult intact male dogs were enrolled. Blood samples were collected to measure CPSE concentrations before (T0), immediately after (T1), and 24 h post (T2) ejaculation. Data were compared within and between groups by ANOVA (p < 0.05). Dogs were divided in two equal groups: A (healthy: CPSE ≤ 52.3 ng/mL at T0) and B (suspected for prostatic disorders: CPSE > 52.3 ng/mL or diagnosed with symptoms of prostatic diseases: CPSE > 90 ng/mL). CPSE was shown to be statistically higher in group B than A at any time point. In both groups, CPSE showed a significant increase at T1, and no significant differences between T0 and T2. This study demonstrates a definite effect of ejaculation on CPSE concentration. Twenty-four hours post-ejaculation, CPSE returns to basal values. Such physiological effects of ejaculation should be considered when planning analyses of CPSE concentrations, i.e., by respecting a proper sexual rest