Abundance of Anti-Muellerian hormone in cat ovaries and correlation of its plasma concentration with animal age, weight and stage of the estrous cycle.

In female animals of different species, Anti-Müllerian hormone (AMH) is produced by follicular granulosa cells and has been associated with the ovarian follicle pool. Because concentration of AMH in plasma of ovary-intact female cats is apparently more variable than previously assumed, we have analysed AMH concentration in blood of cats (n = 93) presented for routine ovariectomy and assessed ovarian histology and AMH protein expression in the surgically removed ovaries. We hypothesised that AMH is synthesized only in preantral and small antral follicles and that plasma AMH concentration reflects the antral follicle count (AFC). Corpora lutea were detected in 35% of the female cats, whereas plasma progesterone concentration was ≥1 ng/mL in 57% of the cats. Follicular cysts were present in 15 cats (16%). Positive immunostaining for AMH protein was detected in close to all primordial and antral follicles, ovarian cysts, 70% of corpora lutea and 28% of atretic follicles. Concentration of AMH in plasma averaged 6.8 ± 0.5 ng/mL (range 1.3-21.7 ng/mL). The AFC increased with increasing AMH concentration with a moderate positive correlation between AFC and AMH (r = 0.286, p < 0.01). Plasma AMH concentration was not affected by season or cats' age, weight, stage of the estrous cycle and presence of follicular cysts. In conclusion, AMH protein is expressed in all endocrine structures of the cat ovary. While AMH is a marker for the presence of ovarian tissue, its usefulness to assess ovarian function in individual female cats is of limited value.

Age-related changes in circulating INSL3 concentrations and their associations with ovarian conditions in Japanese Black beef cattle.

Information on circulating levels of insulin-like peptide 3 (INSL3) in female domesticated animals is limited, as their concentrations are significantly lower than in males. The objectives of the present study were to 1) develop a sandwich time-resolved fluorescence immunoassay (TRFIA) with higher detectability to measure blood INSL3 concentrations in female cattle, 2) determine INSL3 concentrations in female cattle among age groups and reproductive conditions, and 3) explore associations between INSL3 levels and ultrasonographic ovarian measurements. Blood was collected repeatedly from Japanese Black beef female calves (n = 12; 0－8 mo), heifers (n = 10; 10－26 mo), and cows (n = 20; 27－200 mo). Blood was taken from the cows (n = 13) at follicular, post-ovulatory, and luteal phases, and from cows with follicular cysts (n = 12). Ultrasonography of ovaries was conducted in the calves (n = 12) and the cows without ovarian diseases (n = 9). The ovarian area, as well as the number and diameters of antral follicles ≥ 2 mm, were determined in each ovary. The proposed method detected a difference in plasma INSL3 between calves (0.01 ng/mL) and heifers (0.18 ng/mL). However, the conventional assay showed similar levels for calves and heifers (1.82 vs 2.07 ng/mL). Plasma INSL3 and testosterone concentrations increased from calves to heifers (P < 0.0001), but only INSL3 rose from heifers to cows (P < 0.0001). INSL3 and testosterone concentrations did not change across the estrus cycle in cows, and the levels of both hormones in follicular cystic cows did not differ from those in the follicular phase. Ovarian area, maximal and average follicular diameters, and total volume of all follicles per animal were higher in cows than calves (P < 0.001). Plasma INSL3 concentrations correlated positively with the total volumes of all follicles in calves (P < 0.05) and cows (P < 0.05), whereas testosterone concentrations did not correlate with ovarian follicular measurements. In conclusion, plasma INSL3 concentrations measured by the proposed sandwich TRFIA showed a clear increase from female calves to cows in beef cattle. These results suggest that circulating levels of INSL3, but not of testosterone, are associated with the total volume of all antral follicles in both ovaries per animal in female cattle.

A conscious minimally invasive approach to remove follicular cysts and infundibular keratinizing acanthomas in five dogs.

Follicular cysts and infundibular keratinizing acanthomas are common benign cutaneous lesions in dogs. Current treatment options include surgical excision under general anaesthesia, cryotherapy, carbon dioxide laser and retinoids, each with potential disadvantages. We describe a conscious, minimally invasive, surgical excision procedure with high success rate and no complications in five dogs.

Les kystes folliculaires et les acanthomes kératinisant infundibulaires sont des lésions cutanées bénignes fréquentes chez le chien. Les options de traitement actuelles comprennent l'exérèse chirurgicale sous anesthésie générale, la cryothérapie, le laser au dioxyde de carbone et les rétinoïdes, chacun présentant des inconvénients potentiels. Nous décrivons une procédure d'exérèse chirurgicale consciente, peu invasive, avec un taux de réussite élevé et aucune complication chez cinq chiens.

Los quistes foliculares y los acantomas queratinizantes infundibulares son lesiones cutáneas benignas comunes en perros. Las opciones de tratamiento actuales incluyen la escisión quirúrgica bajo anestesia general, crioterapia, láser de dióxido de carbono y retinoides, cada uno con posibles desventajas. Describimos un procedimiento de escisión quirúrgica consciente, mínimamente invasivo, con una alta tasa de éxito y sin complicaciones en cinco perros.

Cistos foliculares e acantomas infundibulares queratinizantes são lesões cutâneas benignas em cães. As opções terapêuticas existentes atualmente são excisão cirúrgica sob anestesia geral, crioterapia, laser de dióxido de carbono e retinoides, cada um com desvantagens potenciais. Nós descrevemos aqui um procedimento de excisão cirúrgica minimamente invasivo, consciente, com um grande potencial de sucesso e sem complicações em cinco cães.

Locally extensive follicular hamartomas with concurrent follicular cysts and dermoid cysts on the head of a dog.

A 5-month-old, male intact Australian shepherd dog was presented with nodular lesions affecting the dorsal head. A locally diffuse form of follicular hamartomas, with concurrent dermoid cysts and follicular cysts, was diagnosed by histopathological examination. Complete surgical excision of the affected skin and closure with a subdermal plexus advancement flap led to long-term resolution.

Un berger australien mâle intact âgé de 5 mois a été présenté avec des lésions nodulaires affectant la face dorsale de la tête. Une forme localement diffuse d'hamartomes folliculaires, avec des kystes dermoïdes et des kystes folliculaires concomitants, a été diagnostiquée par examen histopathologique. L'éxérèse chirurgicale complète de la peau atteinte et la fermeture avec un lambeau d'avancement du plexus sous-cutané ont conduit à une résolution à long terme.

Un perro pastor australiano macho entero de 5 meses de edad se presentó con lesiones nodulares que afectaban el dorso de la cabeza. Mediante examen histopatológico se diagnosticó una forma localmente difusa de hamartomas foliculares, con quistes dermoides y quistes foliculares concurrentes. La escisión quirúrgica completa de la piel afectada y el cierre con una zona avanzada en colgajo del plexo subdérmico condujo a una resolución a largo plazo.

Topical Imiquimod Therapy for Localized Solar Dermatitis in a Dog.

Chronic sun exposure in dogs can result in clinical changes in the skin referred to as solar or actinic dermatitis. A 5-year-old, neutered male, American Bulldog presented with localized unilateral erythematous macules, plaques, alopecia, comedones and hemorrhagic bullae involving the non-pigmented skin on left lateral ventral flank area. The presence of hyperplastic epidermis with keratinocyte dysplasia, superficial dermal elastosis, and multiple follicular cysts with occasional rupture (furunculosis) on histology, together with history and characteristic skin lesions was consistent solar dermatitis/actinic keratosis. Skin scrapings were negative and treatment for secondary pyoderma was initiated with oral clindamycin for 8 weeks. After 2 months of antibiotic therapy, the hemorrhagic bullae resolved; however, erythematous solar/actinic skin lesions with induration and comedones persisted. Topical application of imiquimod 5% cream 3 times weekly for 8 weeks resulted in the resolution of erythema, but some of the non-inflamed comedones remained. Staining for elastin and Ki67 revealed keratinocyte hyperplasia in hair follicle infundibulums and alterations in the elastic fibers around follicles, which may lead to closure and formation of follicular cysts. Imiquimod has been long suggested as a treatment option for solar dermatitis, but this is the first known case report detailing its efficacy in dogs.

A review on inflammation and angiogenesis as key mechanisms involved in the pathogenesis of bovine cystic ovarian disease.

Cystic ovarian disease (COD) is an important cause of reproductive failure in dairy cattle. The main aim of this review is to discuss some aspects related to inflammation and angiogenesis that seem to be involved in the development of follicular cysts in domestic animals, with special emphasis on the bovine species, in an attempt to elucidate the relationship between these two processes in the early stages of persistence and in the development of bovine COD. We describe the changes in the expression of cytokines and angiogenic factors that seem to generate disturbances in the intraovarian component underlying the aberrant persistence of follicular cysts. Results show that pro-inflammatory and anti-inflammatory cytokines behave as regulators of angiogenesis through direct and indirect effects, like overexpression of pro-angiogenic factors, particularly in bovine ovarian cells from follicular cysts and persistent follicles. We conclude that, in dairy cattle, an imbalance in the expression of cytokines and pro-angiogenic growth factors related to ovulation and the processes associated with it would contribute to follicular persistence and to the recurrent appearance of COD.

The expression and localization of selected matrix metalloproteinases (MMP-2, -7 and -9) and their tissue inhibitors (TIMP-2 and -3) in follicular cysts of sows.

Matrix metalloproteinases (MMPs) are a family of enzymes that degrade extracellular matrix (ECM) molecules, playing a vital role in tissue remodeling under physiological and pathological conditions. Their expression and/or activity are regulated by specific tissue inhibitors of MMPs named TIMPs. Recently, an imbalance in the MMP/TIMP system has been found in human and bovine ovarian cysts, but its role in porcine cyst pathogenesis is unknown. This study examined mRNA expression, protein abundance and localization for selected members of the MMP/TIMP system in follicular cysts of sows. Based on histological analysis, we have assessed follicular (FC) and follicular lutein (FLC) cysts with preovulatory follicles (PF) used as a control. Regarding the pattern of MMP expression, increased MMP2, MMP7 and MMP9 mRNA levels were observed in FLC. Furthermore, both pro- and active forms of MMP-2 and MMP-9 proteins were more abundant in FLC. In FC, the abundance of latent and active forms of MMP-9 and the active form of MMP-2 were greater when compared with PF. In relation to TIMPs, TIMP-2 mRNA and protein expression were increased in FLC, whereas TIMP-3 was up-regulated in both FC and FLC only at the protein level. Using immunofluorescence, MMP-2, MMP-7, TIMP-2 and TIMP-3 were detected in granulosa and theca compartments of FC and within the entire luteinized wall of FLC. Notably, MMP-9 occurred weakly in the granulosa layer of FC, but abundantly in the theca compartment of FC and in the luteinized FLC. Taken together, our findings indicate altered expression of the MMP/TIMP system, suggestive of increased ECM degradation, in sow follicular cysts. These components may be involved in the pathogenesis of porcine ovarian cysts through the ECM remodeling.

Foreign body reaction to ruptured follicular cysts in dogs.

BACKGROUND: Cutaneous cysts are common in dogs, and surgical resection is the recommended treatment. However, additional therapy may be required for ruptured follicular cysts with severe cutaneous complications. CASE PRESENTATION: A 3-year-old neutered male Samoyed was presented with multifocal masses on the forelimbs. A 5-year-old neutered female Maltese was also presented with multiple masses and ruptured lesions, which were ulcerative and painful, around the parotid and submandibular glands. The lesions were examined cytologically. In addition, bacterial and fungal cultures and histopathologic examination were performed. Cutaneous multifocal nodules in the Samoyed could not be diagnosed via cytological examination or bacterial/fungal culture. Histopathology revealed numerous follicular cysts with multiple pyogranulomas of various sizes, some of which contained central keratin debris. In the Maltese, cytologic examination revealed central keratins or enucleated ghost cells in the intact cysts and few keratinized squamous cells mixed with neutrophils, mucus and metachromatic cells in the ruptured cysts. Histopathologic examination revealed severely dilated follicular cysts. Oral steroid and cyclosporine therapy resulted in marked improvement in the aseptic pyogranulomas after 2 weeks in formal case and combined with a surgery for residual cysts in latter case. CONCLUSIONS: We have reported two canine cases of ruptured follicular cysts causing foreign body-like aseptic pyogranulomas around cutaneous tissues and their successful management with pharmacological therapy and surgery.

Association of endometritis and ovarian follicular cyst with mastitis in dairy cows.

The occurrence of multiple metabolic and inflammatory diseases in dairy cows is higher during the periparturient period, which may be triggered by bacterial components, but not a viable bacterium. This study aimed to determine the association of endometritis and ovarian follicular cyst (OFC) with mastitis in dairy cows. Ninety-eight Holstein dairy cows were clinically examined for endometritis and OFC approximately 30-50 days after calving. Blood and milk samples were collected for the determination of milk somatic cell count (SCC); milk interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNFα), and interleukin-8 (IL-8) concentrations; and plasma haptoglobin (Hp) and lipopolysaccharide-binding protein (LBP) concentrations. Of the 98 dairy cows included in this study, 12 were diagnosed with endometritis and 37 cows were identified as OFC-positive, whereas the remaining 49 cows were healthy (without endometritis or OFC). The average and maximum SCCs and plasma Hp and LBP concentrations were not significantly different between the healthy cows and those with endometritis or OFC. However, when the maximum SCC was classified as <300, 300-1,000, or >1,000 × 103 cells/ml, the percentage of cows with the maximum SCC <300 × 103 cells/ml was significantly lower in the endometritis and OFC-positive groups than in the healthy group. These results suggested that cows with endometritis and OFC during the postpartum period exhibit high SCC, indicating that some bacterial components can be transferred between organs.

Altered vitamin D metabolic system in follicular cysts of sows.

This study aimed to examine 25OHD3 concentration in the fluid of follicular and follicular lutein cysts of sows in comparison with preovulatory follicles as well as immunolocalize vitamin D metabolic enzymes (CYP27B1 and CYP24A1) and determine their protein abundances in the cyst wall. We have shown for the first time that 25OHD3 level in the fluid of both cyst types was significantly lower than in preovulatory follicles. Furthermore, we have demonstrated CYP27B1 and CYP24A1 protein immunolocalization and abundance in follicular and follicular lutein cysts. The abundance of protein for both metabolic enzymes was decreased in ovarian cysts when compared to preovulatory follicles. We propose that altered VD metabolism in ovarian cyst might associate with their formation in sows.

Efficacy of ovulation synchronization with timed artificial insemination in treatment of follicular cysts in dairy cows.

The efficacy of five ovulation-synchronization protocols with FTAI in treatment of cows diagnosed with follicular cysts was investigated in a nonrandomized study in a single herd. Cows identified with follicular structures >25 mm on two subsequent ultrasonographic (USG) examinations 7-10 days apart (n = 552) were assigned to one of the five treatment regimens on the day of the second USG examination. Treatment regimens were Ovsynch (GnRH-7d-PGF2α-56 h-GnRH-16 h-FTAI), New-CIDR (as Ovsynch with a new CIDR insert being fitted between days 0 and 7), Reused-CIDR (as New-CIDR but the CIDR insert was previously used for 7 days in another cow), G-New CIDR (Pre-GnRH on day 0 with the New-CIDR regimen being initiated 7 days later), and RG-Ovsynch (Pre-GnRH on day 0 and then every 7 days until detection of a luteal structure upon USG examination; at that point the Ovsynch was initiated). A subset of cows was subjected to ovarian USG examination at the time of PGF2α administration, at insemination, and 8-10 days post-insemination. Progesterone-releasing ability of new and reused CIDR inserts were evaluated in cows diagnosed with severe ovarian inactivity (n = 16). The data were analyzed using logistic regression with pregnancy per AI on days 30 (P/Al 30) and 70 (P/AI 70) post-insemination were included as outcome measures. Compared with Ovsynch, RG-Ovsynch improved the P/AI 30 (OR = 2.6, P = 0.03) and the P/AI 70 (OR = 2.5, P = 0.05). New-CIDR and G-New CIDR were associated with non-significant increase in P/AI 30 (OR = 2.1, P = 0.09 and OR = 2.3, P = 0.07, respectively) and P/AI 70 (OR = 2.01, P = 0.09 and OR = 2.2, P = 0.09, respectively). Reused-CIDR was not associated with improvement in P/AI (P = 0.93 and 0.79 for P/AI 30 and P/AI 70, respectively). RG-Ovsynch had a longer diagnosis-to-FTAI interval (median 24, IQR 17,31). The dominant ovarian structures and the presence of a cyst or a luteal structure at PGF2α administration or at insemination were not associated with P/AI. The new and re-used CIDR inserts produced comparable concentrations of serum progesterone 3 h, 3 days and 7 days post CIDR insertion. In conclusion, the RG-Ovsynch improved the P/AI whereas the New-CIDR and the G-New CIDR regimens tended to increase the P/AI compared with Ovsynch. Marginal differences in P/AI between RG-Ovsynch, New-CIDR, and G-New-CIDR together with prolonged diagnosis-to-FTAI in RG-Ovsynch should be considered if to evaluate the economic value of these regimens.

Association of RBP-4 gene polymorphisms with follicular cysts in large white sows.

Follicular cysts, which is a common infertility disease, can cause financial losses in pig breeding programmes. The pathogenesis and mechanisms of the formation of follicular cysts are not understood clearly. In our previous study, the concentration of retinol-binding protein 4 (RBP-4) in the follicular fluid (FF) of the ovary with follicular cysts was found to be significantly higher than that of normal ovary, thereby suggesting that RBP-4 may be a candidate biomarker for porcine follicular cysts. To study the association of RBP-4 and follicular cysts further, we detected the polymorphisms of the RBP-4 gene and the presence of follicular cysts by PCR-Restriction fragment length polymorphism (RFLP) assay. In this study, we screened the mutations of RBP-4 gene in 79 sows with follicular cysts and 100 normal sows without cysts. Results showed that +249-63G>C polymorphisms were significantly associated with follicular cysts, and sows with CC genotype in RBP-4 gene had a high risk of developing follicular cysts. Hence, our findings further proved that RBP-4 may be a novel biomarker for follicular cysts, which may be valuable for the diagnosis of follicular cysts and molecular breeding of pigs.

Levels of selected cytokines and acute-phase proteins in the serum of dairy cows with cystic ovarian disease and those in follicular and luteal phases of normal ovarian cycle.

The aim of the study was to evaluate the concentration of selected cytokines (tumor necrosis factor [TNFα], interleukin [IL]-6, IL-10) and acute-phase proteins (APPs; haptoglobin [Hp] and serum amyloid A [SAA]) in the serum of cows with follicular and luteal ovarian cysts and in those in the follicular and luteal phases of a physiological ovarian cycle. The study was conducted on 24 Holstein-Friesian (HF) cows: eight cows in the follicular phase of the cycle; the same cows after 10 days when they were in the luteal phase; eight cows with follicular cysts; and eight cows with luteal cysts. Levels of progesterone, 17ß-estradiol, TNF-α, IL-6, IL-10, SAA, and Hp were evaluated in the serum of all examined cows. The concentration of the assessed parameters in the serum was determined using commercially available bovine ELISA kits that were specific for each parameter. The results obtained then showed that values of TNF-α and IL-6 were highest in cows with luteal cysts (p < .001), compared to those with other ovarian structures. The highest level of IL-10 was recorded in cows with two types of ovarian cysts. Furthermore, high values of SAA and Hp were found in cows with two types of cysts; however, these values were higher in cows with follicular cysts. The present study shows that an assessment of the levels of cytokines (pro- and anti-inflammatory) and APPs in the serum may be important in the investigation of the processes underlying the formation and differentiation of ovarian cysts in dairy cows.

Successful control of disseminated follicular cysts in a dog with low dose isotretinoin.

A case of disseminated cysts in a dog is described. Histopathological examination revealed the presence of follicular infundibular cysts, which were treated with isotretinoin at a dose of 2 mg/kg body weight (BW), q24h for 1 week, followed by a dose of 1 mg/kg BW for 3 months. Symptoms resolved after this course of treatment.

Contrôle réussi de kystes folliculaires disséminés chez un chien à l'aide d'une faible dose d' isotrétinoïne . Nous décrivons un cas de kystes disséminés chez un chien. L'examen histopathologique a révélé la présence de kystes infundibuliformes folliculaires qui ont été traités à l'aide de l'isotrétinoïne à une dose de 2 mg/kg poids corporel (PC), q24h pendant 1 semaine, suivie d'une dose de 1 mg/kg PC pendant 3 mois. Les symptômes se sont résorbés après ce régime de traitement.(Traduit par Isabelle Vallières).

Kisspeptin and bovine follicular cysts.

The aim of this study was to investigate kisspeptin (Kp) and sexual hormone blood concentrations in healthy dairy cows (H) and in cows diagnosed with ovarian follicular cysts (FC). Forty dairy cows were enrolled in this study and divided in 2 groups of 20 subjects each. All cows underwent blood collection on the day of diagnosis and on that of heat detection, respectively. Kisspeptin 10, estradiol, and progesterone blood concentrations were assessed. All the parameters were higher in Group FC than in Group H. These results suggest that the higher steroid levels found may have determined the increase in Kp secretion in the cystic cows. This, in turn, may have directly or indirectly stimulated the sustained Luteinizing Hormone (LH) release, known to occur in dairy cystic disease.

Ovarian lesions in 44 rabbits (Oryctolagus cuniculus).

The clinical and pathological records of 44 domestic, female rabbits with an age ranging from 6-124 months (median age: 63.5 month) were assessed retrospectively for ovarian lesions. Included were all rabbits that underwent an ovariohysterectomy with a subsequent pathological examination of the genital tract between March 1997 and June 2016. Pathological examination revealed ovarian lesions in 12 of the 44 rabbits including follicular cysts (n=7), cystic rete ovarii (n=3), widespread ovarian necrosis with dystrophic calcification (n=2), ovarian adenoma (n=1). Clinical examination including radiographs only suggested ovarian disorders in two cases of ovarian necrosis with dystrophic calcification and in two cases of cystic rete ovarii. Clinical significance was only conclusive in a case of cystic rete ovarii.

Association between liver failure and hepatic UDP-glucuronosyltransferase activity in dairy cows with follicular cysts.

Uridine 5'-diphospho-glucuronosyltransferase (UGT) liver activity was measured using estradiol-17ß as a substrate in dairy cows with follicular cysts. The activity was significantly lower than that in dairy cows with normal estrous cycles (P<0.01). Liver disorders, such as fatty liver and hepatitis, were observed in half cows with follicular cysts, and liver UGT activity was lower than that in cows with normal estrus cycles. In addition, the liver UGT activity was significantly lower in dairy cows with follicular cysts without liver disorders than in dairy cows with normal estrous cycles. Therefore, the cows were divided into those with low, middle and high liver UGT activities, and liver disorder complication rates were investigated. The complication rate was significantly higher in the low- (78.1%) than in the middle- (22.2%) and high-level (8.3%) groups, suggesting that liver disorders are closely associated with the development of follicular cysts in dairy cows and that steroid hormone metabolism is delayed because of reduced liver UGT activity, resulting in follicular cyst formation. We conclude that reduced estradiol-17ß glucuronidation in the liver and liver disorders are associated with follicular cyst occurrence in dairy cows.

Application of an innovative technique for unilateral ovariectomy in dairy cows.

This report describes an innovative unilateral ovariectomy technique for the surgical treatment of follicular cysts in dairy cows. Five cows, with a diagnosis of follicular cysts, were submitted to unilateral ovariectomy via a paralumbar fossa approach, in standing position. The linear cutter proved useful for clamping the ovary, stapling the vessels and cutting in one stroke, thus reducing surgery times. After unilateral ovariectomy, all cows underwent heat 7-30 days after surgery, and three cows became pregnant. All cows showed increased milk production. The results of this case report showed that flank ovariectomy, performed in this way, is an efficient, low-risk technique for surgical treatment of follicular cysts in dairy cows.

A new insertion/deletion fragment polymorphism of inhibin-α gene associated with follicular cysts in Large White sows.

Ovarian follicular cysts are anovulatory follicular structures that lead to infertility. Hormones play key roles in the formation and persistence of cysts. Inhibins are heterodimeric gonadal glycoprotein hormones that belong to the transforming growth factor-ß superfamily. These hormones suppress the secretion of follicle-stimulating hormone. In this report, partial fragment of inhibin-α (INHA) subunit gene of Large White pig was detected from the genomic DNA by polymerase chain reaction. The sequence showed a 283 bp fragment insertion/deletion (I/D) polymorphism in INHA subunit gene. A total of 49 Large White sows with cystic follicles and 152 normal sows were screened for this polymorphism. The relationship of INHA I/D polymorphisms with follicular cysts was investigated. The distribution of I/D was significantly different between cystic and normal sows, thereby suggesting that the INHA subunit gene might be a potential biological marker for breeding programs in pig.