Expression pattern of germ cell markers in cryptorchid stallion testes.

Cryptorchidism affects spermatogenesis and testis development, often resulting in stallion subfertility/infertility. This study aims to identify the specific germ cells impacted by cryptorchism in stallions. In a previous study, we found that PGP9.5 and VASA are molecular markers expressed in different germ cells within stallions. Herein, we assessed the heat stress-induced response of spermatogonial stem cells (SSCs) in the seminiferous tubules (ST) of cryptorchid stallion testes (CST) and normal stallion testes (NST). This goal was accomplished by comparing PGP9.5 and VASA expression patterns through reverse transcription quantitative PCR and immunofluorescence assays. We also compared the cross-sectional ST area between groups. Six post-pubertal Thoroughbred unilateral cryptorchid stallions were used. The relative abundance of the mRNA transcripts of PGP9.5 and VASA was significantly upregulated in the NST group than in the CST group. Additionally, the cross-sectional ST area and localization of PGP9.5 and VASA in germ cells were significantly higher in the NST group than in the CST group. Regarding Leydig cells, PGP9.5 staining was observed in both groups. Spermatogonia, primary spermatocytes and secondary spermatocytes were immunostained with VASA in the NST group, while immunostaining was only observed in spermatogonia in the CST group. These results indicate long-term exposure to heat stress conditions, such as cryptorchidism, directly impacts germ cell proliferation and differentiation, leading to impaired spermatogenesis and compromised fertility in stallions.

Heme oxygenase 1 (HO1) regulates autophagy and apoptosis via the PI3K/AKT/mTOR signaling pathway of yak Sertoli cells.

Successful male reproduction depends on healthy testes. Autophagy has been confirmed to be active during many cellular events associated with the testes. It is not only crucial for testicular spermatogenesis but is also an essential regulatory mechanism for Sertoli cell (SCs) ectoplasmic specialization integrity and normal function of the blood-testis-barrier. Hypoxic stress induces oxidative damage, apoptosis, and autophagy, negatively affecting the male reproductive system. Cryptorchidism is a common condition associated with infertility. Recent studies have demonstrated that hypoxia-induced miRNAs and their transcription factors are highly expressed in the testicular tissue of infertile patients. Heme oxygenase 1 (HO1) is a heat-shock protein family member associated with cellular antioxidant defense and anti-apoptotic functions. The present study found that the HO1 mRNA and protein are up-regulated in yak cryptorchidism compared to normal testes. Next, we investigated the expression of HO1 in the SCs exposed to hypoxic stress and characterized the expression of key molecules involved in autophagy and apoptosis. The results showed that hypoxic stress induced the upregulation of autophagy of SCs. The down-regulation of HO1 using siRNA increases autophagy and decreases apoptosis, while the over-expression of HO1 attenuates autophagy and increases apoptosis. Furthermore, HO1 regulates autophagy and apoptosis via the PI3K/AKT/mTOR signaling pathway. These results will be helpful for further understanding the regulatory mechanisms of HO1 in yak cryptorchidism.

A massive alteration of gene expression in undescended testicles of dogs and the association of KAT6A variants with cryptorchidism.

Cryptorchidism is the most common form of disorder of sex development in male dogs, but its hereditary predisposition is poorly elucidated. The gonadal transcriptome of nine unilaterally cryptorchid dogs and seven control dogs was analyzed using RNA-seq. Comparison between the scrotal and inguinal gonads of unilateral cryptorchid dogs revealed 8,028 differentially expressed genes (DEGs) (3,377 up-regulated and 4,651 down-regulated). A similar number of DEGs (7,619) was found by comparing the undescended testicles with the descended testicles of the control dogs. The methylation status of the selected DEGs was also analyzed, with three out of nine studied DEGs showing altered patterns. Bioinformatic analysis of the cDNA sequences revealed 20,366 SNP variants, six of which showed significant differences in allelic counts between cryptorchid and control dogs. Validation studies in larger cohorts of cryptorchid (n = 122) and control (n = 173) dogs showed that the TT genotype (rs850666472, p.Ala1230Val) and the AA genotype in 3'UTR (16:23716202G>A) in KATA6, responsible for acetylation of lysine 9 in histone H3, are associated with cryptorchidism (P = 0.0383). Both the transcript level of KAT6A and H3K9 acetylation were lower in undescended testes, and additionally, the acetylation depended on the genotypes in exon 17 and the 3'UTR. Our study showed that the massive alteration of the transcriptome in undescended testicles is not caused by germinal DNA variants in DEG regulatory sequences but is partly associated with an aberrant DNA methylation and H3K9 acetylation patterns. Moreover, variants of KAT6A can be considered markers associated with the risk of this disorder.

Testicular expression of heat SHOCK proteins 60, 70, and 90 in cryptorchid horses.

Heat shock proteins are the most evolutionarily conserved protein families induced by stressors including hyperthermia. In the context of pathologies of the male reproductive tract, cryptorchidism is the most common genital defect that compromises the reproductive potential of the male because it induces an increase in intratesticular temperature. In equine species, cryptorchidism affects almost 9 % of newborns and few studies have been carried out on the molecular aspects of the retained testis. In this study, the expression pattern of HSP60, 70, and 90 in abdominal and inguinal testes, in their contralateral descended normally testes, and in testes of normal horses were investigated by Western blot and immunohistochemistry. The histomorphological investigation of retained and scrotal testes was also investigated. The seminiferous epithelium of the retained testes showed a vacuolized appearance and displayed a completely blocked spermatogenesis for lacking meiotic and spermiogenetic cells. On the contrary, the contralateral scrotal testes did not show morphological damage and the seminiferous epithelium displayed all phases of the spermatogenetic cycle as in the normal testes. The morphology of Leydig cells was not affected by the cryptorchid state. Western blot and immunohistochemistry evidenced that equine testis (both scrotal and retained) expresses the three investigated HSPs. More in detail, the Western blot evidenced that HSP70 is the more expressed chaperone and that together with HSP90 it is highly expressed in the retained gonad (P < 0.05). The immunohistochemistry revealed the presence of the three HSPs in the spermatogonia of normal and cryptorchid testes. Spermatogonia of retained testes showed the lowest expression of HSP60 and the highest expression of HSP90. Spermatocytes, spermatids of scrotal testes, and the Sertoli cells of retained and scrotal testes did not display HSP60 whereas expressed HSP70 and HSP90. These two proteins were also localized in the nucleus of the premeiotic cells. The Leydig cells displayed the three HSPs with the higher immunostaining of HSP70 and 90 in the cryptorchid testes. The results indicate that the heat stress condition occurring in the cryptorchid testis influences the expression of HSPs.

Half-life of serum anti-Müllerian hormone and changes after gonadectomy in adult female and male dogs with normal and abnormal gonads.

Anti-Müllerian hormone (AMH) is a biomarker for the presence of gonadal tissue. Changes in serum AMH after gonadectomy are not well established, and its serum half-life is unknown in dogs. We measured serum AMH with a validated electro-chemiluminescent immunoassay in adult female (n = 12) and male (n = 7) dogs with normal gonads, as well as in dogs with gonadal pathology (ovarian remnant syndrome, ORS n = 3, testicular tumor [Leydig cell, Sertoli cell, seminoma] n = 3, unilateral abdominal cryptorchid n = 4) on the day of gonadectomy (D0), and on D3, D7, D14 (females and males), and D21, D28 (males only). Males had higher AMH concentrations than females independent of gonadal status (P < 0.001). Dogs with ORS had lower initial AMH (0.45 ± 0.43 ng/ml) than bitches with normal gonads (1.16 ± 0.44 ng/ml; P = 0.027). Cryptorchid dogs had higher initial concentrations (80.57 ± 52.81 ng/ml) than males with normal gonads (7.92 ± 2.45 ng/ml; P = 0.004), and those with testicular tumors (18.63 ± 5.04 ng/ml) were intermediate (P ≥ 0.250). AMH decreased over time (P ≤ 0.012) and was 0.01-0.04 ng/ml by D14 in females and 0.02-0.12 ng/ml by D28 in males. Serum half-life in the whole study population was 2.85 ± 0.51 days and did not differ between groups. In conclusion, serum AMH can differentiate between intact and gonadectomized status of adult dogs by 14 days after ovario(hyster)ectomy in females and by 28 days after surgical castration in males.

Histological study of Bactrian camel cryptorchidism and expression of immunoglobulin λ light chain in the testicular and epididymis of cryptorchid Bactrian camel.

Lambda light chains (λ-LCs) are frequently responsible for triggering the activation of inflammatory factors in autoimmune disorders, and an increase in their levels will cause various pathological changes in serum. The aim of this study was to determine the histological differences between the epididymis and testis of normal and cryptorchid Bactrian camels and the differences in λ-LC expression in the epididymis and testis of normal and cryptorchid Bactrian camels. Haematoxylin and eosin (H&E) staining was used to examine the pathological changes in cryptorchidism. The gene and protein levels of λ-LC were determined using RT-qPCR and western blot. The distribution of λ-LCs was assessed by immunohistochemistry and immunofluorescence. Compared with that in normal Bactrian camels, the diameter of the epididymal lumen and the thickness of the epithelium were decreased in the epididymis of cryptorchidic animals. Additionally, no sperm was detected in the cavity of the cryptorchidic epididymis. Meanwhile, the expression of λ-LC was significantly increased in the cryptorchidic epididymis at both the mRNA and protein levels (p < .05). The highest protein expression of λ-LC was found in epididymal epithelial halo cells and testicular Sertoli cells. These findings suggested that the structural changes observed in the epididymal epithelium of cryptorchidic camels affect its secretory and absorptive functions. Additionally, the high level of λ-LC expression recorded in halo cells suggested that these cells play an important role in epithelial immunity in cryptorchidic Bactrian camels. Furthermore, the high λ-LC expression levels detected in normal testicular Sertoli cells indicated that λ-LCs may be involved in spermatogenesis. The results of this study provide clues for an in-depth study of immunological sterility in cryptorchidic Bactrian camels.

Single-port laparoscopic-assisted abdominal cryptorchidectomy in 14 dogs.

OBJECTIVE: To describe a modified laparoscopic-assisted cryptorchidectomy technique in dogs using a single-port endoscope and evaluate clinical outcome in abdominal cryptorchid dogs that underwent the procedure. STUDY DESIGN: Prospective case series. ANIMALS: A total of 14 client-owned dogs (19 abdominal cryptorchid testes). METHODS: Dogs scheduled for laparoscopic cryptorchidectomy between January 2019 and April 2022 were enrolled in the study. The dogs underwent single-port laparoscopic-assisted cryptorchidectomy (SP-LAC) performed by a single surgeon using a 10-mm single-port endoscope placed in the midline immediately cranial to the prepuce. The abdominal testis was endoscopically located and grasped, the cannula was retracted, the capnoperitoneum was reversed to allow exteriorization of the testis, and the spermatic cord was ligated extracorporeally. RESULTS: Median age was 13 months (range, 7-29 months) and median bodyweight was 23.0 kg (range, 2.2-55.0 kg). Nine of 14 dogs had unilateral abdominal cryptorchidism (7 right-sided and 2 left-sided) and 5/14 dogs had bilateral abdominal cryptorchidism. Median surgical time for unilateral abdominal cryptorchidectomy was 17 min (range, 14-21 min) and for bilateral abdominal cryptorchidectomy 27 min (range, 23-55 min). Ten dogs had additional surgical procedures performed concurrently with SP-LAC. One major intraoperative complication (testicular artery hemorrhage) occurred that required emergency conversion and two minor entry-related complications were observed. CONCLUSION: The SP-LAC procedure enabled removal of abdominal testes and was associated with a low morbidity. CLINICAL SIGNIFICANCE: The SP-LAC procedure can be performed by a single surgeon and represents a less invasive alternative to multi-port laparoscopic-assisted or single-port multi-access laparoscopic cryptorchidectomy techniques.

Laparoscopic cryptorchidectomy using a resorbable self-locking loop device in dorsally recumbent horses.

OBJECTIVE: To report the use and feasibility of a self-locking resorbable loop device for cryptorchidectomy. STUDY DESIGN: Clinical prospective study. POPULATION: Twenty horses. METHODS: Horses suspected to have an abdominal testicle/s after admission work-up were enrolled. Horses were anesthetized in dorsal recumbency and a standard laparoscopic technique was performed. The looped device was inserted into the abdomen, glided around the testis/cord and tightened. Then, the spermatic cord was transected prior testis removal. Surgical procedure details and remarks, perioperative complications and total surgical time were recorded. Short- (>3 weeks) and long-term (>6 months) follow-ups were obtained by telephone questionnaire. RESULTS: Median total surgical time was 67 min (range: 43-189 min) and significantly shortened after the first four horses. The loop device was easily glided around 13/20 abdominal testes and required more time and technical skills around larger testes (≥3 years). Excellent intraoperative hemostasis was achieved in 17 horses. Three horses demonstrated mild intraoperative bleeding that required retightening, device replacement or adding a second device, respectively. Three horses developed mild postoperative hemoabdomen identified ultrasonographically and were successfully managed medically. Follow-up revealed no significant complications related to the procedure. One horse was euthanized for colic 4 months after surgery and one died of hemolytic shock 17 months postoperatively. CONCLUSION: This device represents another method to perform equine cryptorchidectomy that requires minimal training and laparoscopic expertise but demands knowledge of the device and application to prevent complications. CLINICAL SIGNIFICANCE: Laparoscopic cryptorchidectomy using this device is an alternative technique for horses <3 years.

Abnormalities of the Scrotum and Testes.

Abnormalities of the bovine scrotum and testes are an important cause of infertility. Proper evaluation of the male reproductive system is a critical first step in screening for such abnormalities. Excessive periscrotal fat, cutaneous scrotal defects, and unilateral scrotal swelling are common deformities that warrant further investigation. Early diagnosis and surgical intervention are often needed to restore reproductive soundness. This article reviews these conditions and provides therapeutic modalities.

MicroRNA as epigenetic regulators of canine cryptorchidism.

Cryptorchidism, the failed descent of one or both testes into the scrotum, is a common developmental disorder in male dogs. Cryptorchidism may affect canine fertility, reducing the quality of the semen, and may promote spermatic cord torsion and onset of neoplasia. MicroRNAs (miRNAs) are epigenetic regulators of gene expression and their dysregulation is associated with disorders of spermatogenesis and testis neoplasia. The present study aimed at investigating the expression of miRNAs in formalin-fixed, paraffin-embedded (FFPE) canine retained testes and testes affected by seminoma, and at integrating miRNAs to their target genes. Forty testicular FFPE specimens from 30 dogs were included - 10 scrotal and 10 contralateral retained from 10 unilateral cryptorchid dogs; 10 tumoral testes affected by seminoma from non-cryptorchid dogs; 10 scrotal normal testes from non-cryptorchid dogs included as the control. The expression level of three miRNAs, namely miR-302c-3p, miR-302a-3p, and miR-371-3p, associated with testicular disorders, were quantified using RT-qPCR. The comparative analysis demonstrated that the level of miR-302a-3p and miR-371a-3p were quantifiable exclusively in control testes. The expression level of miR-302c-3p was higher in the control than in the other groups; its expression decreased in retained testes compared to scrotal testes and testes with seminoma. Gene Ontology analysis pointed out that these miRNAs may be involved in the modulation of estrogen and thyroid hormone signaling pathways. In conclusion, this study demonstrated that miRNAs are dysregulated in canine cryptorchid and seminoma-affected testes compared to control tissues, confirming the pivotal role of miRNAs in cryptorchidism.

Urethrorectal fistula ligation and transection in a dog with a history of unilateral abdominal cryptorchidism and misdiagnosed anal gland disease.

A 10-month-old unilaterally cryptorchid male labradoodle dog was presented to a specialty surgery referral center with a reported history of persistent anal sac leakage that was unresponsive to conservative management. The dog underwent castration and left anal sacculectomy but was witnessed urinating from the anus postoperatively. A contrast urethrogram was performed, and a urethrorectal fistula located between the distal portion of the pelvic urethra and the caudal aspect of the rectum was diagnosed. One week later, the dog underwent ligation and transection of the fistula. Surgery was successful and the dog recovered unremarkably with complete resolution of clinical signs. A contrast urethrogram 1 mo after surgery confirmed the successful ligation and transection of the fistula. Key clinical message: To the authors' knowledge, this is the first clinical report of urethrorectal fistula ligation and transection in a dog with unilateral abdominal cryptorchidism and misdiagnosed anal sac disease. Our findings underscore the importance of a thorough physical examination, history taking, and clinical workup for appropriate diagnosis and treatment of multiple congenital abnormalities.

Ligature et transection de la fistule urétrorectale chez un chien ayant des antécédents de cryptorchidie abdominale unilatérale et de maladie des glandes anales mal diagnostiquée. Un chien Labradoodle mâle unilatéralement cryptorchide âgé de 10 mois a été présenté à un centre de référence en chirurgie spécialisée avec des antécédents signalés de fuite persistante du sac anal qui ne répondait pas à une prise en charge conservatrice. Le chien a subi une castration et une sacculectomie anale gauche, mais a été vu en train d'uriner par l'anus après l'opération. Un urétrogramme de contraste a été réalisé et une fistule urétrorectale située entre la partie distale de l'urètre pelvien et la face caudale du rectum a été diagnostiquée. Une semaine plus tard, le chien a subi une ligature et une section de la fistule. La chirurgie a été un succès et le chien s'est rétabli sans particularité avec une résolution complète des signes cliniques. Un urétrogramme de contraste 1 mois après la chirurgie a confirmé la réussite de la ligature et de la section de la fistule.Message clinique clé :À la connaissance des auteurs, il s'agit du premier rapport clinique de ligature et de transection de fistule urétrorectale chez un chien atteint de cryptorchidie abdominale unilatérale et d'une maladie du sac anal mal diagnostiquée. Nos résultats soulignent l'importance d'un examen physique approfondi, d'une anamnèse et d'un bilan clinique pour un diagnostic et un traitement appropriés des anomalies congénitales multiples.(Traduit par Dr Serge Messier).

Anti-Müllerian hormone levels in serum and testes of male dogs: relations with neuter status and bilateral abdominal cryptorchidism.

Anti-Müllerian hormone (AMH) analysis has contributed to new information in the reproductive endocrinology of domestic animals, due to clinically available diagnostic tools. An accurate and rapid diagnostic method to distinguish between neutered and bilateral abdominal cryptorchid dogs is needed in veterinary practice. Therefore, this study uses an enzyme-linked immunosorbent assay to evaluate the clinical relevance of AMH analysis in peripheral blood as a diagnostic tool for dogs with suspected bilateral abdominal cryptorchidism. The possible alteration of the AMH localization in testicular tissue caused by this pathologic condition was also investigated using immunohistochemistry. Male dogs were divided into three groups of healthy intact (n = 14), healthy castrated (n = 14), and bilateral abdominal cryptorchid (n = 14) dogs. The results demonstrated a higher level of serum AMH in the cryptorchid group compared to the intact group (P < 0.01), while serum AMH levels of all castrated dogs were below the limit of detection (<0.05 ng/mL). Moreover, the percentage of positive AMH immunostaining of the intact group was less than that of the cryptorchid group (P < 0.01). A significantly positive correlation was found between serum AMH concentration and localization in testicular tissues (r = 0.93, P < 0.01). Our findings suggest that AMH levels in the peripheral blood could be used as an alternative and rapid screening method for detecting dogs with abdominal cryptorchidism.

Efficiency of immunocastration with an anti-gonadotropin-releasing hormone vaccine on cryptorchid bulls.

Cryptorchid bulls have low economic value owing to the effects of masculinization. Moreover, surgical removal of an ectopic testis is difficult in certain clinical cases. Recently, immunocastration has garnered popularity as a nonsurgical castration method in pig farming; however, the effects of immunocastration on cryptorchid bulls are yet to be yet. Herein, we investigated endocrine changes due to immunocastration in cryptorchid bulls and studied its effectiveness. This study included 13 Holstein bulls diagnosed with cryptorchidism and classified into two groups based on pubertal period: <8 months of age (pregroup) and ≥8 months of age (postgroup). Antigonadotropin-releasing hormone (GnRH) vaccine was used for immunocastration, and two vaccine doses were administered. Blood testosterone and anti-Müllerian hormone (AMH) levels were measured and analyzed for endocrine evaluation. The testosterone levels significantly decreased following the start of immunocastration in both groups, thereby confirming the efficacy of antiGnRH vaccination in cryptorchid bulls. The AMH levels significantly increased in the pregroup with two antiGnRH vaccination, suggesting a compensatory response via the neutralization of GnRH antibodies. The AMH levels did not significantly change in the postgroup, indicating the partial suppression of AMH secretion in Sertoli cells during sexual maturation and failure of Sertoli cell maturation. Thus, we successfully restrained the serum testosterone levels in cryptorchid bulls using antiGnRH vaccine. The testosterone levels are a useful indicator of the immunocastration effect on cryptorchid bulls. Hereafter, a vaccine program that can sustain the castration effect on cryptorchid bulls is necessary.

Whole-Exome Sequencing Indicated New Candidate Genes Associated with Unilateral Cryptorchidism in Pigs.

INTRODUCTION: Cryptorchidism is a hereditary anomaly characterized by the incomplete descent of one or both testicles to the scrotum. One of the challenges of this anomaly is that the retained testicle maintains its endocrine function. As a consequence, cryptorchid animals produce hormone-tainted meat in comparison to castrated animals and are likely to be more aggressive. Cryptorchidism can lead to reduced animal welfare outcomes and cause economic losses. Identifying genetic markers for cryptorchidism is an essential step toward mitigating these negative outcomes and may facilitate genome manipulation to reduce the occurrence of cryptorchidism. Attempts to identify such markers have used genome-wide association studies. Using whole-exome sequencing, we aimed to identify single nucleotide polymorphisms (SNPs) in the coding regions of cryptorchid pigs and to characterize functional pathways concerning these SNPs. METHODS: DNA was extracted and sequenced from 5 healthy and 5 cryptorchid animals from the Landrace breed, using the Illumina HiSeq 2500 platform. Data were pre-processed using the SeqyClean tool and further mapped against the swine reference genome (Sus scrofa 11.1) using BWA software. GATK was used to identify polymorphisms (SNPs and InDels), which were annotated using the VEP tool. Network prediction and gene ontology enrichment analysis were conducted using the Cytoscape platform, and STRING software was used for visualization. RESULTS: A total of 63 SNPs were identified across the genes PIGB, CCPG1, COMMD9, LDLRAD3, TRIM44, MYLPF, SEPTIN, ZNF48, TIA1, FAIM2, KRT18, FBP1, FBP2, CTSL, DAPK1, DHX8, GPR179, DEPDC1B, ENSSSCG00000049573, ENSSSCG00000016384, ENSSSCG00000022657, ENSSSCG00000038825, and ENSSSCG00000001229. Using pathway enrichment analyses and network prospection, we have identified the following significant adjusted p value threshold of 0.001 involved with the biological function pathways of estrogen signaling, cytoskeleton organization, and the pentose phosphate pathway. CONCLUSION: Our data suggest the involvement of new SNPs and genes in developing cryptorchidism in pigs. However, further studies are needed to validate our results in a larger cohort population. Variations in the GPR179 gene, with implications at the protein level, may be associated with the appearance of this anomaly in the swine. Finally, we are showing that the estrogen signaling pathway may be involved in the pathophysiological mechanisms of this congenital anomaly as previously reported in GWAS.

Update on canine anorchia: A review.

Abnormalities of the external genitals are an important issue in dog breeding because of the unfavourable qualities and characteristics of breeds, resulting in consistent economic losses. Despite their significance, little scientific attention has been given to these problems. Although there are several reviews on cryptorchidism in dogs, none have described anorchia. Testicular agenesis is a rare reproductive disorder with a congenital origin. Moreover, no author has described the diagnostic procedure for making a definitive diagnosis of anorchia in dogs. It is important to have a well-structured diagnostic scheme to help practical veterinarians make a confirmatory diagnosis. This review article aims to provide an update on canine anorchia diagnosis based on the poor research studies published in recent years. We have also contributed to the pathogenesis of this disorder using human medicine studies. Finally, the review includes therapeutic hypotheses that can be expanded in future studies.

Polyorchidism in a cat.

A 9-mo-old, male domestic shorthair cat was presented for castration because of mounting behavior observed by the owner. On physical examination, the cat was bilaterally cryptorchid, but had penile spines. Abdominal exploration through a midline laparotomy revealed 2 pairs of masses. All 4 masses had gross features of testes, and ranged from 7 × 5 × 5 mm to 12 × 6 × 7 mm, with associated epididymal tissue. Histologically, each mass contained seminiferous tubules consistent with testicular tissue, and epididymal tubules, confirming a diagnosis of polyorchidism; deferent ducts were not found. There was no evidence of neoplastic, infectious, or inflammatory disease. Mounting behavior ceased 4 wk post-surgery. Histologic confirmation of more than 2 testes is needed to establish a diagnosis of polyorchidism, a rare congenital anomaly that has been reported infrequently in the veterinary literature; reports have been of animals with triorchidism, with the exception of 1 cat with 4 intraabdominal testes. Our report emphasizes that, although rare, polyorchidism should be considered in cryptorchid cats, or whenever penile spines are present in a previously castrated cat. Our case also highlights the value of checking for penile spines in a bilaterally cryptorchid cat if abdominal ultrasound is not an option to aid in surgical planning.

Usefulness of computed tomography for cryptorchidism in bulls.

Cryptorchidism is defined as the failure of the testis to descend into the scrotal position. Bulls with cryptorchidism have problems in both meat quality and husbandry management; thus, it is greatly important to accurately identify the retained testis and remove it during the early stage. Abdominal computed tomography (CT) was performed under general anesthesia in 34 bulls aged 3-9 months with cryptorchidism. All bulls underwent laparoscopic or incision approach for cryptorchidectomy, and 40 testes were dissected. The detection rates of retained testes were 64.5% in the abdominal cavity and 100% in the subcutaneous inguinal region, and the total detection rate was 72.5%. Furthermore, all cases in this study were suspected to have intra-abdominal cryptorchidism in primary care, but CT revealed that 22.5% of cases had cryptorchidism in the subcutaneous inguinal region. The CT value (mean ± standard deviation) of the retained testes was 20.96 ± 7.54 Hounsfield Unit, and the CT value and size of the retained testes showed a positive weak correlation with bovine age. Therefore, there is the demerit that general anesthesia and a huge device are necessary; nevertheless, CT is suggested to be useful in identifying the location of retained testes and selecting an appropriate surgical approach in bulls with cryptorchidism. Moreover, CT findings suggested that the maturation of the retained testes might depend not on the descending process but on age.

Melatonin through blockade of Hif-1α signaling mediates the anti-fibrosis under hypoxia in canine Sertoli cells.

The hypoxic microenvironment of cryptorchidism is an important factor in the impairment and fibrosis of Sertoli cells which result in blood-testis barrier (BTB) destruction and spermatogenesis loss. Recent studies have shown that melatonin, a well-known pineal hormone exerts beneficial effects against pathological fibrosis in a various of organs. However, it is still unknown whether melatonin can regulate hypoxia-induced fibrosis of Sertoli cells. In this study we evaluate melatonin levels, and its synthesizing enzymes, AANAT and HIOMT expression patterns in canine cryptorchidism and contralateral normal testis. Results show abdominal testes presented low melatonin levels and AANAT and HIOMT expression compared with testes located in the scrotum. Moreover, we established a hypoxia-induced fibrosis model in canine Sertoli cells induced by cobalt chloride (CoCl2) and found that melatonin inhibited the EMT markers expression and ECM production as well as Hif-1α expression of Sertoli cells in a dose-dependent manner. Furthermore, use of Lificiguat (synonyms YC-1, Hif-1α inhibitor) to interfere with the Hif-1α pathway showed a similar effect with melatonin suppression of the fibrosis in Sertoli cells. The results indicate that melatonin supplementation can alleviate the fibrosis process of Sertoli cells caused by hypoxia, which is associated with regulating the inhibition of Hif-1α signaling.

Heat stress and hypoxia inhibit the secretion of androgens and induce epithelial-to-mesenchymal transition associated with activated TGF-ß/Smad signalling in canine cryptorchidism.

Cryptorchidism, as a common congenital disease of canine testes, is mainly caused by factors leading to endocrine abnormalities in testes and infertility in a heat stress and hypoxia microenvironment. Moreover, heat stress and hypoxia, as critical microenvironmental factors, promote epithelial-mesenchymal transition (EMT), which occurs during adult tissue remodelling responses including carcinogenesis and fibrosis and is the main cause of testicular tumours. In this study, we found by haematoxylin-eosin staining that the canine cryptorchid tissue produced a lot of collagen fibres. Also, the quantitative PCR and Western blot results showed that the mRNA and protein levels of the heat stress makers HSP70 and HO-1 and the hypoxia maker HIF-1α are significant higher compared with normal testes. Moreover, we found the expression levels of TGF-ßs and its two receptors TGF-ßRI and TGF-ßRII increased in case of cryptorchidism. From the study in vitro, we found both heat stress and COCl2 mimic hypoxia inhibited the secretion of testosterone (T) and androstenedione (A4) and promoted the expression of the EMT maker α-SMA and vimentin in Leydig cells, and also that heat stress and COCl2 stimulated with the TGF-ß signalling promoted the expression of TGF-ßs and its two type receptors and also the active phosphorylation of Smad2 and Smad3. The use of LY2109761, a receptor inhibitor of TGF-ßs/Smad signalling pathway, was associated with heat stress and COCl2 suppression of androgens' secretion and stimulated EMT in Leydig cells. These findings characterized a novel pathogenesis of cryptorchidism and provided a new idea for therapeutics.

Computed tomography of testicular torsion in a juvenile dog with unilateral cryptorchidism.

A 14-week-old male unilaterally cryptorchid Clumber spaniel was presented for acute lethargy. Physical examination revealed abdominal pain, and a single testis was palpated in the scrotum. Abdominal ultrasound and computed tomography (CT) revealed a poorly vascularized, ovoid structure immediately caudal to the left kidney with scant regional peritoneal effusion. Left intra-abdominal testicular torsion was confirmed at surgery, and routine cryptorchidectomy was performed. The patient recovered uneventfully from anesthesia and surgery. Key clinical message: The most common CT characteristics of testicular torsion were present in this case and correlated well with sonographic findings to allow for rapid, accurate diagnosis and surgical planning of unilateral, non-neoplastic, intra-abdominal cryptorchid testicular torsion in a juvenile dog. Contrast enhanced CT facilitated accurate localization of the undescended testis and evaluation of testicular perfusion and may be a useful alternative to ultrasound for diagnosing testicular torsion, especially in indeterminate cases.

Tomodensitométrie d'une torsion testiculaire chez un chien juvénile atteint de cryptorchidie unilatérale. Un épagneul Clumber avec une cryptorchidie unilatérale âgé de 14 semaines a été présenté pour une léthargie aiguë. L'examen physique a révélé des douleurs abdominales et un seul testicule a été palpé dans le scrotum. L'échographie abdominale et la tomodensitométrie ont révélé une structure ovoïde mal vascularisée immédiatement caudale au rein gauche avec peu d'épanchement péritonéal régional. Une torsion testiculaire intra-abdominale gauche a été confirmée lors de la chirurgie et une cryptorchidectomie de routine a été réalisée. Le patient s'est remis sans incident de l'anesthésie et de la chirurgie.Message clinique clé:Les caractéristiques tomodensitométriques les plus courantes de la torsion testiculaire étaient présentes dans ce cas et bien corrélées avec les résultats échographiques pour permettre un diagnostic rapide et précis et une planification chirurgicale de la torsion testiculaire avec cryptorchidie unilatérale, non néoplasique et intra-abdominale chez un chien juvénile. La tomodensitométrie avec contraste a facilité la localisation précise du testicule non descendu et l'évaluation de la perfusion testiculaire et peut être une alternative utile à l'échographie pour diagnostiquer la torsion testiculaire, en particulier dans les cas indéterminés.(Traduit par Dr Serge Messier).