The role of inflammation and matrix metalloproteinases in equine endometriosis

Equine endometriosis is a multifactorial disease considered to be a major cause of equine infertility. The purpose of this study was to evaluate the reliability of histomorphological grading for biopsy-like samples compared to entire uterine wall samples, to examine the association between the degree of endometriosis with animal age, and to investigate the role of inflammation in endometriosis and the expression of different matrix metalloproteinases in equine endometrium. Histomorphological lesions in 35 uterine samples were examined while comparing biopsy-like samples and entire-wall samples. Seventeen uterine samples were stained with antibodies against MMP-2, MMP-9, MMP-14, and TIMP-2. The morphologic evaluation results of the biopsy-like tissue and entire-wall samples were significantly correlated. Endometriosis in older mares (>12 years of age) was more severe than in young mares (2~4 years of age), confirming the positive correlation between animal age and disease severity, while inflammation was poorly related to the degree of endometriosis. MMP-2 and MMP-14 were detected in stromal cells, while MMP-9 and TIMP-2 were both found in stromal and glandular epithelial cells. There were no significant differences in MMPs expression between the two groups (young vs. old mares). Additional studies on the activity of MMPs could further define the role of these enzymes in equine endometriosis.     

Pleural endometriosis in an aged rhesus macaque (Macaca mulatta): a histopathologic and immunohistochemical study

Endometriosis is defined as the presence of endometrial tissue outside the uterine cavity and is one of the most common reproductive abnormalities encountered in women as well as Old World primates. The majority of endometriosis cases in Old World primates occur within the abdominal cavity, with spread to extraabdominal sites considered to be a rare event. A 19-year-old multiparous female rhesus macaque (Macaca mulatta) presented to necropsy for difficulty breathing and weight loss. Grossly, the animal had marked abdominal endometriosis and severe hemoabdomen and hemothorax, the latter of which was accompanied by marked pleural fibrosis. Histologic examination confirmed the abdominal endometriosis and also revealed numerous uterine glands and stroma embedded within the pleural fibrosis. Rafts of endometrial tissue were present within pulmonary lymphatics and the tracheobronchial lymph nodes. Immunohistochemically, all ectopic endometrial tissue had varying degrees of positive immunoreactivity to cytokeratin, vimentin, progesterone and estrogen receptors, and calretinin but was negative for desmin and carcinoembryonic antigen. Pleural endometriosis is an extremely rare manifestation of endometriosis in nonhuman primates. This case report emphasizes lymphatic spread as a likely mechanism for extrauterine endometriosis.     

Isolation of multiple drug-resistant enteric bacteria from feces of wild Western Lowland Gorilla (Gorilla gorilla gorilla) in Gabon

Prevalence of drug-resistant bacteria in wildlife can reveal the actual level of anthropological burden on the wildlife. In this study, we isolated two multiple drug-resistant strains, GG6-2 and GG6-1-1, from 27 fresh feces of wild western lowland gorillas in Moukalaba-Doudou National Park, Gabon. Isolates were identified as Achromobacter xylosoxidans and Providencia sp., respectively. Minimum inhibitory concentrations of the following 12 drugs—ampicillin (ABPC), cefazolin (CEZ), cefotaxime (CTX), streptomycin (SM), gentamicin (GM), kanamycin (KM), tetracycline (TC), nalidixic acid (NA), ciprofloxacin (CPFX), colistin (CL), chloramphenicol (CP) and trimethoprim (TMP)—were determined. Isolate GG6-2 was resistant to all antimicrobials tested and highly resistant to CTX, SM, TC, NA and TMP. Isolate GG6-1-1 was resistant to ABPC, CEZ, TC, CL, CP and TMP.     

Bilateral hydronephrosis secondary to endometriosis managed by endoscopic ureteral stent placement in a captive Guinea baboon (Papio papio)

Spontaneous endometriosis is an estrogen-dependent, progressive and painful disease that affects a variety of nonhuman primates, including several species of baboons (Papio sp.). This case documents multimodal management of severe endometriosis in a captive female baboon within a zoological institution. An 18-yr-old, intact female Guinea baboon (Papio papio) was found to have an enlarged uterus. Fifteen months post ovariohysterctomy, scarring associated with endometrial tissue resulted in ureteral strictures, bilateral hydronephrosis, and azotemia. Cystoscopic placement of bilateral ureteral stents with fluoroscopy was performed and resulted in short-term clinical improvement. The animal's condition declined and euthanasia was elected 4 mo after ureteral stent placement. Severe endometriosis with secondary inflammation resulting in bilateral hydroureter and hydronephrosis, as well as concurrent cystitis, ureteritis, and pyelonephritis were confirmed at necropsy. Despite possible complications, ureteral stents can be considered a useful therapeutic option in patients with ureteral disease.     

Chorionic villus sampling for sex determination in a western lowland gorilla (Gorilla gorilla gorilla).

Chorionic villus sampling was undertaken on an anesthetized gorilla (Gorilla gorilla gorilla) to determine the fetal sex in the first trimester of pregnancy. The tissue samples were subject to sex determination by polymerase chain reaction, fluorescent in situ hybridization, and cell culture with cytogenetic analysis. Polymerase chain reaction testing was found to be the most accurate and rapid method of sex determination on this tissue sample.     

子宫内膜异位症大鼠模型复制

取Wistar大鼠30只,用手术移植的 方法把自体子宫内膜组织2块分别移植到壁 腹膜与子宫系膜处,建立子宫内膜异位症大 鼠模型。术后4周,再次开腹,观察异位内膜 的存活情况、与周围组织的粘连程度及病理 学变化。结果发现腹膜和子宫系膜处移植内 膜的成活率及病灶的体积差异无显著性(P >0.05),腹膜处病灶与周围组织的粘连程度 显著轻于子宫系膜处病灶(P<0.01),移植 物外观呈囊状,表面有血管,光镜下观察见子 宫内膜细胞、间质细胞、腺体和血管,与正常 在位内膜相比腺体减少。该模型复制方法简 便,成功率高,是研究子宫内膜异位症较理想 的动物模型。     

Cholesterol values in free-ranging gorillas (Gorilla gorilla gorilla and Gorilla beringei) and Bornean orangutans (Pongo pygmaeus).

Cholesterol concentrations in captive gorillas and orangutans vary widely within species and average approximately 244 mg/dl for gorillas and 169 mg/dl for orangutans as published previously. The International Species Inventory System reports higher concentrations of 275 and 199 mg/dl for gorillas and orangutans, respectively. It is unknown whether these values were typical, influenced by captive management, or both. To answer this question, banked serum samples from free-ranging mountain gorillas (Gorilla beringei), western lowland gorillas (Gorilla gorilla gorilla), and Bornean orangutans (Pongo pygmaeus) were analyzed for total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol concentrations. Mountain gorillas did not differ significantly from free-ranging western lowland gorillas in cholesterol, triglyceride, high-density lipoprotein cholesterol, or low-density lipoprotein cholesterol concentrations, indicating mountain gorilla values could be a model for western lowland gorillas. Captive gorilla total cholesterol and low-density lipoprotein cholesterol concentrations were significantly higher (P < 0.05) than in free-ranging groups. Triglyceride concentrations for captive gorillas were significantly higher (P < 0.05) than the male mountain and western lowland gorillas, but they were not significantly different from the female mountain gorillas. Captive orangutan total cholesterol concentrations were only higher (P < 0.05) than the free-ranging female orangutans, whereas captive orangutan low-density lipoprotein cholesterol concentrations were significantly higher (P < 0.05) than both free-ranging male and female orangutans. Calculated and measured low-density lipoprotein cholesterol concentrations were compared for all free-ranging animals and were significantly different (P < 0.05) for all groups, indicating Friedewald's equation for calculating low-density lipoprotein cholesterol is not appropriate for use with nonfasted apes. The higher total cholesterol and low-density lipoprotein cholesterol concentrations in captive apes may predispose them to cardiovascular disease and might be attributed to diets, limited energy expenditure, and genetics.     

Rectal prolapse in a free-ranging mountain gorilla (Gorilla beringei beringei): clinical presentation and surgical management.

A juvenile female mountain gorilla (Gorilla beringei beringei) of the Mubare tourist group in Bwindi Impenetrable National Park, Uganda, developed a severe, complete rectal prolapse that did not spontaneously resolve. Eight months prior, a juvenile female mountain gorilla of the Mubare group developed a mild, complete rectal prolapse that resolved spontaneously within 24 hr. Field guides reported that spontaneously resolving prolapses had been seen previously in two other juveniles, one of which was from the Mubare group. The tissue became increasingly necrotic and maggot infested over the course of 1 wk. Surgical intervention involved amputation of the affected rectal tissues and suturing the viable portion to the anal sphincter muscle with simple interrupted absorbable sutures. The surgery was performed in the field in accordance with Uganda Wildlife Authority policies. Antibiotics and anthelmintics were administered systemically, and the gorilla returned to the group. The gorilla appeared to recover fully after 3 wk. Histology of the resected rectal tissue confirmed intense inflammation and necrosis with myiasis but did not reveal an underlying etiology.     

Long-bone fractures in llamas and alpacas: 28 cases (1998–2008)

Treatment and outcome of camelids with long-bone fractures are described. Medical records (1998–2008) of camelids (n = 28) with long-bone fractures were reviewed for signalment, time to presentation, fracture type, method of repair, duration of hospitalization, and post-operative complications. Follow-up information was obtained via telephone interviews with owners. Mean age and weight at presentation were 3.4 years and 56.3 kg, respectively. Twenty-six fractures were treated with internal fixation (n = 11), external fixation (n = 10), combination of internal and external fixation (n = 3), amputation (n = 1), and external fixation followed by amputation (n = 1). Long-term follow-up information was obtained for 19 of the 26 animals. The post-operative complication rate was 23% and owner satisfaction was high. Animals with open fractures were more likely to experience complications. Internal fixation was associated with superior alignment and outcome. Internal fixation techniques should be recommended for camelids.     

The response of adipose tissues to Mycoplasma pulmonis and Sendai virus infection in C57BL/6 and DBA/2 mice

Adipose tissues in mammals are categorized into white and brown adipose tissues in which cellular morphology, cell functions, and tissue distribution are different. White adipose tissue (WAT) plays a major role in energy reservation, while brown adipose tissue (BAT) mainly relates to the thermoregulation of the body. One interesting function of adipose tissue is the response to the infection, especially the pathogens that cause pneumonia. We have previously reported that DBA/2 (D2) mice are susceptible to pathogens causing pneumonia, Mycoplasma (M.) pulmonis and Sendai virus (SeV), whereas C57BL/6 (B6) mice are resistant to them. Furthermore, morphological alteration of mediastinal fat tissue (MFT) was seen after infection of M. pulmonis in D2 mice but not in B6 mice. In this study, we aimed to exhibit the difference in adipose tissue response in other areas, including interscapular brown adipose tissue (iBAT), inguinal white adipose tissue (ingWAT), and perigonadal WAT (perigoWAT) between resistant strain, B6 and susceptible strain, D2 after challenging them with M. pulmonis and SeV. Compared with B6 mice, D2 mice showed an increase in fat-associated lymphoid cluster in MFT, an increase in BAT in both iBAT and ingWAT after M. pulmonis and SeV infection. The results of this study indicate that pneumonia caused by M. pulmonis and SeV infection induces browning of adipocyte, suggesting that BAT plays a role in pathogen infection and inflammation.     

ENDOMETRIOSIS AND A PARAOVARIAN CYST IN A RHESUS MACAQUE

We describe endometriosis in an aged rhesus macaque. There was a large mass and a related paraovarian cyst, typical of the disease. Endometriosis is a common finding in nonhuman primate. In this report, we also review the pathophysiology of the disease and summarize the historical and more recent relevant literature. Given the frequency of endometriosis in the rhesus monkey and the long-life spans (15-30 years) of nonhuman primates in captivity, endometriosis should be suspected in animals displaying the earliest signs of the disease: anorexia, dysmenorrhea, menorrhagia, irregular menstrual cycles, or infertility. Despite recent advances in the diagnosis and therapeutic strategies for endometriosis in women, the disease remains a significant cause of morbidity and ultimately, a cause of mortality, in the older nonhuman primate.     

Fibrous osteodystrophy in a dromedary camel

A 6-y-old female dromedary camel (Camelus dromedarius L.) was presented for assessment of firm, bilateral swellings rostral and ventral to the eyes. Serum biochemistry revealed hyperglycemia (28.5 mmol/L), hypocalcemia (1.27 mmol/L), hyperphosphatemia (3.39 mmol/L), hypoproteinemia (total protein 50 g/L), and hypoalbuminemia (20 g/L). Based on the poor prognosis associated with the presumptive diagnosis of fibrous osteodystrophy, the camel was euthanized. Gross postmortem findings revealed expanded fibrous tissue replacing the maxilla and mandible, and bilaterally prominent parathyroid glands. Histology of the maxilla revealed proliferative loose fibrous tissue with widely scattered, regularly spaced, small spicules of mineralized bone. The parathyroid glands were prominent bilaterally; the internal and external parathyroid glands were composed of plump cells with abundant pale basophilic cytoplasm and open nuclei. The pathologic findings were consistent with the antemortem diagnosis of fibrous osteodystrophy. The camel’s diet, which was not specifically balanced for a camel, included grass hay, sweet feed, and alfalfa pellets. The camel’s caregivers reported feeding her treats of cookies. A feed analysis was not available. The biochemistry abnormalities and clinical and postmortem findings, along with a diet that was not balanced for a camel, are consistent with a diagnosis of nutritional secondary hyperparathyroidism.     

Vaccination of pigs with the S48 strain of Toxoplasma gondii – safer meat for human consumption

As clinical toxoplasmosis is not considered a problem in pigs, the main reason to implement a control strategy against Toxoplasma gondii (T. gondii) in this species is to reduce the establishment of T. gondii tissue cysts in pork, consequently reducing the risk of the parasite entering the human food chain. Consumption of T. gondii tissue cysts from raw or undercooked meat is one of the main sources of human infection, with infected pork being considered a high risk. This study incorporates a mouse bioassay with molecular detection of T. gondii DNA to study the effectiveness of vaccination (incomplete S48 strain) in its ability to reduce tissue cyst burden in pigs, following oocyst (M4 strain) challenge. Results from the mouse bioassay show that 100% of mice which had received porcine tissues from vaccinated and challenged pigs survived compared with 51.1% of mice which received tissues from non-vaccinated and challenged pigs. The presence (or absence) of T. gondii DNA from individual mouse brains also confirmed these results. This indicates a reduction in viable T. gondii tissue cysts within tissues from pigs which have been previously vaccinated with the S48 strain. In addition, the study demonstrated that the main predilection sites for the parasite were found to be brain and highly vascular muscles (such as tongue, diaphragm, heart and masseter) of pigs, while meat cuts used as human food such as chop, loin, left tricep and left semitendinosus, had a lower burden of T. gondii tissue cysts. These promising results highlight the potential of S48 strain tachyzoites for reducing the number of T. gondii tissues cysts in pork and thus improving food safety.     

Echocardiographic parameters of captive western lowland gorillas (Gorilla gorilla gorilla)

A total of 163 echocardiographic studies on western lowland gorillas (Gorilla gorilla gorilla) were submitted for evaluation; 140 from 99 animals were suitable for analysis. Of these, 81 studies (42 studies from 35 males ranging in age from 11-41+ yr and 39 studies from 31 females ranging in age from 11-41+ yr) are reported here. Three studies from 3 females and 56 studies from 30 males were excluded from this report due to cardiac abnormalities. Cardiac parameters measured were aortic root (Ao Rt) diameter and left atrial (L atrium) size. Left ventricular (LV) measurements included left ventricular internal diameter in systole (LVID(s)) and diastole (LVID(d)) as well as diastolic septal (IVS) and posterior wall thickness (LVPW). Values considered to be normal in females > 11 yr of age were: Ao Rt < 3.5 cm, L atrium < 4.0 cm, LVID(d) < 5.0 cm, IVS < 1.4 cm, LVPW < 1.4 cm, and ejection fraction (EF) > 60%. The data from male gorillas show a separation in animals based on three cardiac parameters: systolic function, LV cavity size, and LV wall thickness. Male gorillas > 11 yr of age fall into two groups; unaffected and affected. Unaffected animals are defined as those with no echocardiographic abnormalities and a consistent Ao Rt of < 4.0 cm, LVID(d) of < 6.0 cm, IVS and LVPW of <1.5 cm, and an EF of > 58%. The affected group consisted of male gorillas that exhibited changes in echocardiographic parameters representing the presence of cardiovascular disease. The results determined in this database, gathered from data collected from 1999-2009, suggest a sex-based difference between males and females with predominantly males demonstrating evidence of cardiac disease. The most striking finding seen in this study is that of progressive LV hypertrophy and depressed LV EF in affected adult male gorillas.     

Potassium permanganate elicits a shift of the external fish microbiome and increases host susceptibility to columnaris disease

The external microbiome of fish is thought to benefit the host by hindering the invasion of opportunistic pathogens and/or stimulating the immune system. Disruption of those microbial communities could increase susceptibility to diseases. Traditional aquaculture practices include the use of potent surface-acting disinfectants such as potassium permanganate (PP, KMnO₄) to treat external infections. This study evaluated the effect of PP on the external microbiome of channel catfish and investigated if dysbiosis leads to an increase in disease susceptibility. Columnaris disease, caused by Flavobacterium columnare, was used as disease model. Four treatments were compared in the study: (I) negative control (not treated with PP nor challenged with F. columnare), (II) treated but not challenged, (III) not treated but challenged, and (IV) treated and challenged. Ribosomal intergenic spacer analysis (RISA) and pyrosequencing were used to analyze changes in the external microbiome during the experiment. Exposure to PP significantly disturbed the external microbiomes and increased catfish mortality following the experimental challenge. Analysis of similarities of RISA profiles showed statistically significant changes in the skin and gill microbiomes based on treatment and sampling time. Characterization of the microbiomes using 16S rRNA gene pyrosequencing confirmed the disruption of the skin microbiome by PP at different phylogenetic levels. Loss of diversity occurred during the study, even in the control group, but was more noticeable in fish subjected to PP than in those challenged with F. columnare. Fish treated with PP and challenged with the pathogen exhibited the least diverse microbiome at the end of the study.     

Porcine circovirus type 2 (PCV2) vaccination reduces PCV2 in a PCV2 and Salmonella enterica serovar Choleraesuis coinfection model

We previously reported that prior porcine circovirus type 2 (PCV2) infection potentiates the severity of clinical signs, lung lesions, and fecal shedding and tissue dissemination of Salmonella enterica serovar Choleraesuis in infected pigs. Here, we evaluated whether PCV2 vaccination is effective in reducing fecal shedding and tissue dissemination of S. Choleraesuis and improving clinical signs associated with PCV2 and S. Choleraesuis infection in 15 Cesarean-derived, colostrum-deprived pigs randomly assigned to 3 groups (n = 5/group). The vaccinated and co-infected (VAC-COINF) group received 2 ml of a commercial PCV2 vaccine at age 3 weeks. The VAC-COINF and co-infected (COINF) groups were inoculated intranasally with PCV2 and S. Choleraesuis at 5 and 7 weeks of age, respectively. The CONTROL group pigs received a similar volume of PBS for sham-vaccination and sham-inoculation. PCV2 vaccination clearly reduced PCV2 DNA load in the serum and postmortem tissue samples and decreased PCV2 antigen levels in tissue samples of the VAC-COINF group. After S. Choleraesuis infection, the incidence of several clinical signs increased in the VAC-COINF group compared to that in the COINF group. The microscopic lung lesions and weight gain, fecal shedding and tissue dissemination of S. Choleraesuis except in the spleen were not significantly different in the VAC-COINF and COINF groups. Thus, PCV2 vaccination reduced PCV2 in the S. Choleraesuis and PCV2 coinfection model and the effects on S. Choleraesuis were minimal.     

Identification of Helicobacter DNA in feline pancreas,liver, stomach,and duodenum: Comparison between findings in fresh and formalin-fixed paraffin-embedded tissue samples

The clinical significance of Helicobacter spp. in feline digestive organs needs to be evaluated and formalin-fixed and paraffin-embedded (FFPE) tissue samples provide an invaluable source for molecular studies. In this study, we performed a PCR assay to investigate the presence of Helicobacter DNA in digestive organs from seven cats and compared this occurrence in fresh and formalin-fixed and paraffin-embedded (FFPE) tissue samples from the same organs. The present study identified Helicobacter DNA in the pancreas, liver, stomach, and duodenum in fresh tissue samples but only in the stomach in FFPE samples. To our knowledge this is the first time that Helicobacter DNA have been identified in the feline pancreas. This study indicates that it is important to be aware of differences between results when analyzing FFPE samples compared to fresh tissue samples, especially regarding longer DNA fragments (>200 bp (base pairs)).     

Excretory–secretory antigens: A suitable candidate for immunization against ocular toxoplasmosis in a murine model

Toxoplasmosis, responsible for ocular impairment, is caused by Toxoplasma gondii. We investigated the effect of Toxoplasma excretory–secretory antigens (ESA) on parasite load and distribution in the eye tissue of a murine model. Case and control groups were immunized with ESA and PBS, respectively. Two weeks after the second immunization, the mice were challenged intraperitoneally with virulent RH strain of Toxoplasma; eye tissue samples of both groups were collected daily (days 1, 2, 3, and the last day before death). Parasite load was determined using real-time quantitative PCR targeted at the B1 gene. Compared to the control group, infected mice that received ESA vaccine presented a considerable decrease in parasite load in the eye tissue, demonstrating the effect of ESA on parasite load and distribution. Diminution of parasite load in mouse eye tissue indicated that ESA might help control disease-related complications and could be a valuable immunization candidate against ocular toxoplasmosis.     

Experimental Study of Bone Lengthening in Dogs by Means of Backscattered Scanning Electron Microscopy

Objective— To describe the morphology of calcified tissues involved in distraction osteogenesis (DO) by means of backscattered scanning electron microscopy (BS-SEM).
Study Design— Experimental study.
Animals— Adult female Beagle dogs (n=12).
Methods— Nonsimultaneous and bilateral transverse mid-diaphyseal osteotomies performed in tibiae were stabilized and distracted by a Type Ia external skeletal fixation device. After a latency period of 5 days, distraction was applied at a rate of 0.5 mm every 12 hours for 10 days. Then, the external fixator was maintained in a static mode during the consolidation period until bone healing or euthanasia at 1, 2, 3, 4, 6, 8, 10, 12, 14, and 18 weeks after operations, whichever came first. Distracted regions were isolated and their structure was examined by BS-SEM.
Results— Calcified chondroid tissue was prominent during distraction and calcified cartilaginous tissue during consolidation; both tissues were successively replaced by woven, lamellar, and osteonal bone.
Conclusions— In osteotomized tibia, chondroid tissue is the main component of the mineralization front during distraction, calcified cartilaginous tissue during consolidation, and then both tissues are replaced by woven, lamellar, and osteonal bone. The ossification mechanism of distraction callus is transchondroidal.
Clinical Relevance— BS-SEM is an effective technique for studying progression of bone healing during DO. The presence of chondroid tissue during DO explains why callus mineralization occurs more rapidly during distraction than during static stabilization.