An 8-year-old 532-kg (1,170-lb) primiparous American Paint Horse mare was referred to the J. T. V... more An 8-year-old 532-kg (1,170-lb) primiparous American Paint Horse mare was referred to the J. T. Vaughn Large Animal Teaching Hospital at the Auburn University College of Veterinary Medicine for management of dystocia of 10 hours’ duration. The mare was at 337 days of gestation and had no prior history of illness. The owner observed rupture of the chorioallantois at approximately 9 AM on the day of referral, which was followed by obvious abdominal contractions during the subsequent 8 hours. At 5 PM, the referring veterinarian examined the mare and suggested referral; the mare received xylazine hydrochloride (0.3 mg/kg [0.14 mg/lb], IV) and flunixin meglumine (1.1 mg/kg [0.5 mg/lb], IV) at 6 PM prior to transport. The mare arrived at the university veterinary teaching hospital at 8 PM. At the time of arrival, the mare was quiet, alert, and responsive. Vital signs and results of routine physical examination (excluding examination of the reproductive tract) were within anticipated limits. A jugular venous blood sample was obtained and tested, which yielded a PCV of 48% (reference range, 32% to 48%) and total solids concentration of 6.4 g/dL (reference range, 5.6 to 7.6 g/dL). No fetal membranes or fetal tissues protruded from the vulva. Manual examination of the vaginal canal revealed a fetus in a cranial-longitudinal presentation and dorsosacral position, with bilateral extension of the thoracic limbs. A domed structure, approximately 25 cm in diameter, was identified at the cranial aspect of the pelvic inlet; this structure was presumed to be the head of the fetus. The fetal muzzle was retained in the abdominal cavity ventral and right lateral to the brim of the mare’s pelvis (ie, ventrolateral flexion of the head and neck). The fetus was presumed to be dead because of an absence of reflexive movement when firm digital pressure was applied to the coronary band of either thoracic limb of the fetus and the prolonged duration of the second stage of parturition. Correction of the malposture followed by controlled vaginal delivery was selected as the management option. Preprocedural medications administered to the mare included gentamicin (6.6 mg/kg [3 mg/lb], IV), penicillin G potassium (22,000 U/kg [10,000 U/lb], IV), and flunixin meglumine (1.1 mg/kg, IV). The mare was premedicated with xylazine hydrochloride; anesthesia was induced by IV administration of ketamine and maintained by administration of isoflurane and oxygen via endotracheal tube. The mare was placed in dorsal recumbency in the Trendelenburg position, and the birth canal was lubricated with copious amounts of carboxymethylcellulose. A mechanical hoist was attached to the hind limbs of the mare; it was used to intermittently raise and lower the hindquarters to assist the obstetricians as they attempted to reposition the fetus to correct the ventral flexion of the head and neck. The fetus’ large cranium prevented simultaneous entry of the fetal head and thoracic limbs into the vaginal vault. An attempt to collapse the calvarium by manual application of pressure lateral to the fontanelles was unsuccessful; thus, it was determined that vaginal delivery of the intact fetus was not possible. Cesarean section was recommended to the owner for delivery of the suspected malformed fetus but was declined because of economical constraints. Therefore, fetotomy was pursued as a final attempt to deliver the fetus.
We have investigated GnRH immunization for the treatment of urethral sphincter mechanism incompet... more We have investigated GnRH immunization for the treatment of urethral sphincter mechanism incompetence in ovariectomized bitches. It has been reported that decreasing LH secretion through the use of GnRH agonists temporarily restores continence in some bitches. Therefore, decreasing the circulating LH concentrations by immunizing against GnRH might temporarily maintain continence in incontinent dogs. Sixteen incontinent dogs given phenylpropanolamine (PPA) to control incontinence were recruited for this study. Eleven dogs were immunized against GnRH (novel treatment group) at week 0, and nine dogs were vaccinated again 4 weeks later. Five dogs (standard treatment group) were vaccinated with a placebo twice at 4-week intervals. PPA was discontinued in the novel treatment group 2 weeks after revaccination, and standard-treatment dogs were given PPA for the duration of the study. Blood samples were collected before each treatment and at 6, 8, 10, 12, 16, 20, and 24 weeks and owners recorded episodes of incontinence throughout the study. Ten of the eleven dogs in the novel treatment group experienced side effects as a result of vaccination; two of these dogs experienced more severe side effects after the first vaccination and were withdrawn from the study as a result. Of the nine dogs that completed the vaccination series, four dogs remained continent after PPA was discontinued. For these four dogs, there was no difference in incontinent episodes when they were given PPA versus treatment with the vaccine. All nine novel-treatment dogs developed a GnRH antibody titer and experienced a significant decrease in circulating LH concentrations. In conclusion, GnRH immunization was effective in maintaining continence in four of the nine incontinent ovariectomized dogs, and in these dogs, treatment with the vaccine was comparable with treatment with PPA.
ContentsInvestigations using sustained‐release deslorelin implants at various insertion sites hav... more ContentsInvestigations using sustained‐release deslorelin implants at various insertion sites have shown that this method consistently induces oestrus in anoestus bitches. However, fertility comparisons between implant insertion sites have not been performed. Anestrous beagle bitches received one 2.1 mg deslorelin implant beneath the vestibular mucosa (VM group; n = 6) or in the subcutaneous tissue between the shoulder blades (SubQ group; n = 8). Vestibular implants were removed when serum progesterone concentrations first exceeded 1.5 ng/ml. Vaginal cytologies and blood samples were collected daily and bitches were inseminated during oestrus. Serum progesterone and deslorelin concentrations were measured and pregnancy status was determined using ultrasonography. There were no differences between groups in the intervals between implant administration and the onset of proestrus, the time of the luteinizing hormone surge and the onset of cytologic diestrus. There were also no differences in the number of corpora lutea or foetuses. However, conception rate was significantly lower in the SubQ group. The pregnancy rate did not differ significantly between the VM and SubQ groups [4 out of 6 (66.7%) and 3 out of 8 (37.5%), respectively]. One bitch (16.7%) in the VM group and three bitches (37.5%) in the SubQ group suffered distinct, premature declines in serum progesterone concentrations starting 1–4 weeks after cytologic diestrus. Serum progesterone concentrations did not recover (premature luteal failure), resulting in abortion. Bitches with premature luteal failure in the SubQ group still had serum deslorelin concentrations >100 pg/ml 20 days after implant insertion, suggesting a possible association between prolonged deslorelin release and luteal failure.
Purpose was to investigate if an underlying biological cause exists for the reproductive success ... more Purpose was to investigate if an underlying biological cause exists for the reproductive success in unowned free-roaming (UFR) cats. These cats were presented for surgical sterilization during late summer/early fall. After castration of UFR toms, presence of penile spines was recorded, slides were made from vas deferens secretions for sperm morphology analyses, and testes were hemi-sectioned, formalin-fixed, paraffin-embedded, sectioned, stained with hematoxylin and eosin, and seminiferous tubule diameters were measured. After ovariohysterectomy of UFR queens, total ovarian-uterine weights were recorded. Ovaries were hemi-sectioned, formalin-fixed, paraffin-embedded, sectioned, stained, and follicles were counted. Presence of penile spines did not differ (p = 0.07) between toms 2 - 6 (n = 13) and > 6 months (n = 16). Percentage of normal sperm morphology did not differ (p = 0.39) between toms 2 - 6 (n = 11; 77 ± 11%) and > 6 months (n = 9; 81 ± 13%). Seminiferous tubular diame...
Pyometra is a life-threatening uterine infection that affects 20 - 25% of reproductively intact b... more Pyometra is a life-threatening uterine infection that affects 20 - 25% of reproductively intact bitches. Canine pyometra is characterized by accumulation of leukocytes and exudate within the uterine lumen. Although exact temporal and progressive mechanisms underlying its pathogenesis are not fully understood, a combination of endocrine, structural, inflammatory and bacterial factors are likely involved in most cases. Pyometra develops during luteal phase; progesterone has a key role in establishment of infection with opportunistic bacteria. Cystic endometrial hyperplasia results from proliferation of endometrial glands and is a predisposing factor for development of pyometra. Endometrial proliferation factors exacerbate uterine innate immune responses. Whereas Escherichia coli is the most common organism isolated in up to 90% of cases with a positive culture, genes encoding for adhesins, toxins, and other factors increase virulence of certain strains. A presumptive diagnosis of pyom...
Purpose of this investigation was to determine if luteinizing hormone receptor (LHR) expressionin... more Purpose of this investigation was to determine if luteinizing hormone receptor (LHR) expressionin canine lymphoma was affected by body weight, sex, immunophenotype (B cell, T cell) or tumor stage.Formalin-fixed, paraffin-embedded lymphoma tissue samples from spayed and neutered dogs (n = 40),representing multiple breeds and mixed breeds, were subjected to routine immunohistochemicaltechniques using a polyclonal LHR antibody. Percentage of cells positive for LHR and the stainingintensity (scored 0 - 3) were determined at 400 x magnification. Data were expressed as mean ± standarddeviation and significance was defined as p < 0.05. Differences between sex and tumor phenotype, bodyweight and tumor stage were compared. All tumor samples had cells positive for LHR. However,percentage of cells expressing LHR and its immunostaining intensity varied among individuals. Therewere no significant differences in percentage of LHR positive cells or staining intensity within sex orimmunophenotyp...
Transcriptome gene expression studies of sperm RNA have been utilized in a variety of different s... more Transcriptome gene expression studies of sperm RNA have been utilized in a variety of different species to investigate causes of male infertility. Previous research investigators have optimized cell separation and RNA isolation techniques for each species of interest. To date, no study has been completed for dogs. The objective of this thesis research was to investigate the efficacy of various cell separation techniques in separating sperm cells from somatic cells in the ejaculate to yield a sample of total pure sperm RNA that could be used for a future downstream application. Comparisons were made between the conventional swim-up method and two commercial density gradient centrifugation (DGC) solutions (Bovipure TM and Equipure
An 8-year-old 532-kg (1,170-lb) primiparous American Paint Horse mare was referred to the J. T. V... more An 8-year-old 532-kg (1,170-lb) primiparous American Paint Horse mare was referred to the J. T. Vaughn Large Animal Teaching Hospital at the Auburn University College of Veterinary Medicine for management of dystocia of 10 hours’ duration. The mare was at 337 days of gestation and had no prior history of illness. The owner observed rupture of the chorioallantois at approximately 9 AM on the day of referral, which was followed by obvious abdominal contractions during the subsequent 8 hours. At 5 PM, the referring veterinarian examined the mare and suggested referral; the mare received xylazine hydrochloride (0.3 mg/kg [0.14 mg/lb], IV) and flunixin meglumine (1.1 mg/kg [0.5 mg/lb], IV) at 6 PM prior to transport. The mare arrived at the university veterinary teaching hospital at 8 PM. At the time of arrival, the mare was quiet, alert, and responsive. Vital signs and results of routine physical examination (excluding examination of the reproductive tract) were within anticipated limits. A jugular venous blood sample was obtained and tested, which yielded a PCV of 48% (reference range, 32% to 48%) and total solids concentration of 6.4 g/dL (reference range, 5.6 to 7.6 g/dL). No fetal membranes or fetal tissues protruded from the vulva. Manual examination of the vaginal canal revealed a fetus in a cranial-longitudinal presentation and dorsosacral position, with bilateral extension of the thoracic limbs. A domed structure, approximately 25 cm in diameter, was identified at the cranial aspect of the pelvic inlet; this structure was presumed to be the head of the fetus. The fetal muzzle was retained in the abdominal cavity ventral and right lateral to the brim of the mare’s pelvis (ie, ventrolateral flexion of the head and neck). The fetus was presumed to be dead because of an absence of reflexive movement when firm digital pressure was applied to the coronary band of either thoracic limb of the fetus and the prolonged duration of the second stage of parturition. Correction of the malposture followed by controlled vaginal delivery was selected as the management option. Preprocedural medications administered to the mare included gentamicin (6.6 mg/kg [3 mg/lb], IV), penicillin G potassium (22,000 U/kg [10,000 U/lb], IV), and flunixin meglumine (1.1 mg/kg, IV). The mare was premedicated with xylazine hydrochloride; anesthesia was induced by IV administration of ketamine and maintained by administration of isoflurane and oxygen via endotracheal tube. The mare was placed in dorsal recumbency in the Trendelenburg position, and the birth canal was lubricated with copious amounts of carboxymethylcellulose. A mechanical hoist was attached to the hind limbs of the mare; it was used to intermittently raise and lower the hindquarters to assist the obstetricians as they attempted to reposition the fetus to correct the ventral flexion of the head and neck. The fetus’ large cranium prevented simultaneous entry of the fetal head and thoracic limbs into the vaginal vault. An attempt to collapse the calvarium by manual application of pressure lateral to the fontanelles was unsuccessful; thus, it was determined that vaginal delivery of the intact fetus was not possible. Cesarean section was recommended to the owner for delivery of the suspected malformed fetus but was declined because of economical constraints. Therefore, fetotomy was pursued as a final attempt to deliver the fetus.
We have investigated GnRH immunization for the treatment of urethral sphincter mechanism incompet... more We have investigated GnRH immunization for the treatment of urethral sphincter mechanism incompetence in ovariectomized bitches. It has been reported that decreasing LH secretion through the use of GnRH agonists temporarily restores continence in some bitches. Therefore, decreasing the circulating LH concentrations by immunizing against GnRH might temporarily maintain continence in incontinent dogs. Sixteen incontinent dogs given phenylpropanolamine (PPA) to control incontinence were recruited for this study. Eleven dogs were immunized against GnRH (novel treatment group) at week 0, and nine dogs were vaccinated again 4 weeks later. Five dogs (standard treatment group) were vaccinated with a placebo twice at 4-week intervals. PPA was discontinued in the novel treatment group 2 weeks after revaccination, and standard-treatment dogs were given PPA for the duration of the study. Blood samples were collected before each treatment and at 6, 8, 10, 12, 16, 20, and 24 weeks and owners recorded episodes of incontinence throughout the study. Ten of the eleven dogs in the novel treatment group experienced side effects as a result of vaccination; two of these dogs experienced more severe side effects after the first vaccination and were withdrawn from the study as a result. Of the nine dogs that completed the vaccination series, four dogs remained continent after PPA was discontinued. For these four dogs, there was no difference in incontinent episodes when they were given PPA versus treatment with the vaccine. All nine novel-treatment dogs developed a GnRH antibody titer and experienced a significant decrease in circulating LH concentrations. In conclusion, GnRH immunization was effective in maintaining continence in four of the nine incontinent ovariectomized dogs, and in these dogs, treatment with the vaccine was comparable with treatment with PPA.
ContentsInvestigations using sustained‐release deslorelin implants at various insertion sites hav... more ContentsInvestigations using sustained‐release deslorelin implants at various insertion sites have shown that this method consistently induces oestrus in anoestus bitches. However, fertility comparisons between implant insertion sites have not been performed. Anestrous beagle bitches received one 2.1 mg deslorelin implant beneath the vestibular mucosa (VM group; n = 6) or in the subcutaneous tissue between the shoulder blades (SubQ group; n = 8). Vestibular implants were removed when serum progesterone concentrations first exceeded 1.5 ng/ml. Vaginal cytologies and blood samples were collected daily and bitches were inseminated during oestrus. Serum progesterone and deslorelin concentrations were measured and pregnancy status was determined using ultrasonography. There were no differences between groups in the intervals between implant administration and the onset of proestrus, the time of the luteinizing hormone surge and the onset of cytologic diestrus. There were also no differences in the number of corpora lutea or foetuses. However, conception rate was significantly lower in the SubQ group. The pregnancy rate did not differ significantly between the VM and SubQ groups [4 out of 6 (66.7%) and 3 out of 8 (37.5%), respectively]. One bitch (16.7%) in the VM group and three bitches (37.5%) in the SubQ group suffered distinct, premature declines in serum progesterone concentrations starting 1–4 weeks after cytologic diestrus. Serum progesterone concentrations did not recover (premature luteal failure), resulting in abortion. Bitches with premature luteal failure in the SubQ group still had serum deslorelin concentrations &gt;100 pg/ml 20 days after implant insertion, suggesting a possible association between prolonged deslorelin release and luteal failure.
Purpose was to investigate if an underlying biological cause exists for the reproductive success ... more Purpose was to investigate if an underlying biological cause exists for the reproductive success in unowned free-roaming (UFR) cats. These cats were presented for surgical sterilization during late summer/early fall. After castration of UFR toms, presence of penile spines was recorded, slides were made from vas deferens secretions for sperm morphology analyses, and testes were hemi-sectioned, formalin-fixed, paraffin-embedded, sectioned, stained with hematoxylin and eosin, and seminiferous tubule diameters were measured. After ovariohysterectomy of UFR queens, total ovarian-uterine weights were recorded. Ovaries were hemi-sectioned, formalin-fixed, paraffin-embedded, sectioned, stained, and follicles were counted. Presence of penile spines did not differ (p = 0.07) between toms 2 - 6 (n = 13) and > 6 months (n = 16). Percentage of normal sperm morphology did not differ (p = 0.39) between toms 2 - 6 (n = 11; 77 ± 11%) and > 6 months (n = 9; 81 ± 13%). Seminiferous tubular diame...
Pyometra is a life-threatening uterine infection that affects 20 - 25% of reproductively intact b... more Pyometra is a life-threatening uterine infection that affects 20 - 25% of reproductively intact bitches. Canine pyometra is characterized by accumulation of leukocytes and exudate within the uterine lumen. Although exact temporal and progressive mechanisms underlying its pathogenesis are not fully understood, a combination of endocrine, structural, inflammatory and bacterial factors are likely involved in most cases. Pyometra develops during luteal phase; progesterone has a key role in establishment of infection with opportunistic bacteria. Cystic endometrial hyperplasia results from proliferation of endometrial glands and is a predisposing factor for development of pyometra. Endometrial proliferation factors exacerbate uterine innate immune responses. Whereas Escherichia coli is the most common organism isolated in up to 90% of cases with a positive culture, genes encoding for adhesins, toxins, and other factors increase virulence of certain strains. A presumptive diagnosis of pyom...
Purpose of this investigation was to determine if luteinizing hormone receptor (LHR) expressionin... more Purpose of this investigation was to determine if luteinizing hormone receptor (LHR) expressionin canine lymphoma was affected by body weight, sex, immunophenotype (B cell, T cell) or tumor stage.Formalin-fixed, paraffin-embedded lymphoma tissue samples from spayed and neutered dogs (n = 40),representing multiple breeds and mixed breeds, were subjected to routine immunohistochemicaltechniques using a polyclonal LHR antibody. Percentage of cells positive for LHR and the stainingintensity (scored 0 - 3) were determined at 400 x magnification. Data were expressed as mean ± standarddeviation and significance was defined as p < 0.05. Differences between sex and tumor phenotype, bodyweight and tumor stage were compared. All tumor samples had cells positive for LHR. However,percentage of cells expressing LHR and its immunostaining intensity varied among individuals. Therewere no significant differences in percentage of LHR positive cells or staining intensity within sex orimmunophenotyp...
Transcriptome gene expression studies of sperm RNA have been utilized in a variety of different s... more Transcriptome gene expression studies of sperm RNA have been utilized in a variety of different species to investigate causes of male infertility. Previous research investigators have optimized cell separation and RNA isolation techniques for each species of interest. To date, no study has been completed for dogs. The objective of this thesis research was to investigate the efficacy of various cell separation techniques in separating sperm cells from somatic cells in the ejaculate to yield a sample of total pure sperm RNA that could be used for a future downstream application. Comparisons were made between the conventional swim-up method and two commercial density gradient centrifugation (DGC) solutions (Bovipure TM and Equipure
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