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A controlled clinical trial was carried out to assess the mortality and repellency of a new topical combination of fipronil-permethrin (Effitix® Virbac, Mexico) against Rhodnius prolixus in dogs. Ten medium-size dogs (10-15 kg) with short hair were used. The dogs were exposed to 8 adult triatomines once weekly for 7 weeks. On the control day (D0), the dogs were exposed to the insects without treatment. On D7, the dogs were immediately treated with a spot-on 2.2 ml pipette containing 134 mg of fipronil and 1200 mg permethrin after exposure to the insects. The dose was repeated after 4 weeks following the manufacturer's instructions. Repellency at D0 was, 0 % and the insects had a high blood content. After 12 h post-contact, repellency was 86.3 % and slowly decrease though D21 and D28. On D7, none of the insects survived after 3 h of feeding on the treated dogs. On D14, D35 and D42, all insects died within 12 h post-feeding, whereas no mortality was observed in the control D0 (P < 0.05). The results of this study indicated that administration of the product following the manufacturer's instructions was efficacious at inducing rapid mortality of R. prolixus and therefore could be useful to prevent the transmission of American trypanosomiasis in dogs.

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This study assessed the in vitro anthelmintic (AH) activity of methanol and acetone:water leaf extracts from Annona squamosa, A. muricata and A. reticulata against Haemonchus contortus eggs. The egg hatch test was used to determine the effective concentrations required to inhibit 50% of eggs hatching (EC50). The role of polyphenols on AH activity was measured through bioassays with and without polyvinylpolypyrrolidone (PVPP). Methanolic extracts mainly caused the death of eggs at the morula stage (ovicidal activity). Meanwhile, acetone:water extracts caused egg-hatching failure of developed larvae (larvae failing eclosion (LFE) activity). The lowest EC50 values against H. contortus eggs were observed for the methanolic extracts from A. reticulata and A. muricata (274.2 and 382.9 µg/ml, respectively). From the six extracts evaluated, the methanolic extracts of A. muricata, A. reticulata and A. squamosa showed the highest ovicidal activity, resulting in 98.9%, 92.8% and 95.1% egg mortality, respectively. When the methanolic extract of A. squamosa was incubated with PVPP, its AH activity increased. Similarly, when acetone:water extracts of A. muriata and A. reticulata were incubated with PVPP, their LFE activity increased. Alkaloids were only evident in methanolic extracts, irrespective of PVPP incubation. The presence of acetogenins was not observed. In conclusion, methanolic extracts obtained from leaves of A. muricata, A. reticulata and A. squamosa showed ovicidal activity affecting the morula of H. contortus eggs, with minor LFE activity. Meanwhile, acetone:water extracts showed mostly LFE activity, with a lower proportion of ovicidal activity.

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This study aimed to estimate the seroprevalence and determine the parasite load of Toxoplasma gondii (T. gondii) in the heart and tongue tissues and the semimembranosus/gracilis muscles in a population of Mexican hairless pig (MHP). A cross-sectional study was conducted in 81 MHP, 9-12 weeks of age, from 10 municipalities in Yucatan, Mexico. The prevalence was estimated by the detection of T. gondii IgG antibodies via an indirect ELISA assay. The parasite burden was also determined by testing genomic DNA from sampled tissues using quantitative real-time PCR (qPCR), which amplified a 62bp product from the B1 gene of T. gondii. The seroprevalence was 53% (43/81), ranging from 0 to 100% seropositivity rate depending on the municipalitie of origin. The qPCR allowed detection of T. gondii in 5.3% (9/168) of the tissue samples, with an average of 2.5±2.71 parasites per gram (0.17±0.18 parasite equivalent) for the leg muscle tissue, 0.26±0.39 (0.01±0.02 parasite equivalent) for the heart tissue and 0.31±0.37 (0.021±0.025 parasite equivalent) for the tongue tissue. No significant difference (p>0.05) was observed in the proportion and parasite burden among the different types of tissues evaluated. As shown by the ELISA, a high seroprevalence of T. gondii exists in MHP from the Yucatán Peninsula. The parasite burden found in the tissues showed similar tropism; therefore, the consumption of these tissues involves the same risk of acquiring infection if not properly cooked.

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The objective of this study was to estimate the prevalence and identify risk factors associated with the presence of Toxoplasma gondii in pig-fattening farms from Yucatan, Mexico. A cross-sectional study was conducted with a two-stage sampling. There were 429 pigs sampled from 39 farms randomly selected. Blood samples were collected to obtain DNA and serum. The presence of IgM and IgG antibodies was determined by indirect ELISA. Prevalence was estimated by diagnostic test. Potential risk factors to be included in a marginal logistic regression were tested by chi-square or Fisher. The prevalence of IgM and IgG was 92.5% (397/429) (CI 89.9-95.1%) and 95.8% (411/429) (CI 93.7-97.8%), respectively. Regarding PCR, a prevalence of 50.8% (218/429) (CI 45.9-55.6%) was found. The logistic regression showed an association with herd size and type of feeder (P < 0.05). The risk of a case in farms with ≤400 pigs was 27.9 times higher than in farms with >400 pigs. The manual feeder was a significant protective factor associated with the seropositive against T. gondii. Results indicate a high circulation of T. gondii in pig-fattening farms from Yucatan, finding an increased risk of infection for those farms with less than 400 animals and automatic feeders.

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The epidemiology of Chagas disease is complex. There are different vectors and reservoirs and different clinical manifestations. In order to assess whether the biological behavior of three strains isolated in southeastern Mexico (H4 isolated from human, Z17 isolated from Didelphis sp., and V isolated from T. dimidiata) could be modified during passage through the vector T. dimidiata, the parasitemia curve, the amount of amastigote nests, and mortality of BALB/c infected with blood trypomastigotes of T. cruzi were evaluated. Strains were maintained in continuous passage from mouse to mouse and in animals infected with metacyclic trypomastigotes. The parasitemia curves were significantly different (P < 0.05) between mice to mice and triatoma to mice groups in strains H4 and Z17, and was also observed fewer amastigote nests in cardiac tissue (P < 0.05 strain H4 with higher number versus all groups and Z17 between mice to mice and triatoma to mice) 45 days after inoculation. It is concluded that T. dimidiata influences in modulating the virulence of strains of T. cruzi in the region. Further studies of the intestinal tract of the insect in search for some protein molecules involved in regulating may clarify the virulence of the parasite.

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American trypanosomiasis is an infectious disease of importance for public health and caused by the protozoa Trypanosoma cruzi mainly transmitted by triatomine bugs. The precise role of cats in the peridomestic transmission of T. cruzi and the mechanism by which cats become infected remain uncertain. The objective of this work was to determine the prevalence of T. cruzi infection in domestic cats from an urban area of tropical Mexico by serological and molecular methods and evaluate associated risk factors. A total of 220 domestic cats from Merida Yucatan, Mexico, were studied. Animals older than 3 months were blood sampled. Serum and DNA were obtained. Specific T. cruzi IgG antibodies were detected using a commercial indirect ELISA with an anti-cat antibody HRP labelled. Positive cases were confirmed by Western blot (WB). Polymerase chain reaction (PCR) was also performed using the primers TC1 and TC2. From the 220 cats, 8.6% had antibodies against T. cruzi using ELISA test and later confirmed by WB. In 75 cats (34%), the sequence of ADNk of T. cruzi was amplified. The bad-regular body condition was the only risk factor associated with PCR positive to T.cruzi (P < 0.001). In Mexico, there are no previous epidemiological reports that demonstrate the importance of the cat as a reservoir of T. cruzi. Few individuals were identified with a serological response because they were probably at an early stage of infection or antibodies were not detected because they could be immunocompromised (FIV, FeLV or others). It is necessary to monitor PCR-positive patients and conduct further studies for better understanding of the epidemiology and pathogenesis of Chagas disease in domestic cats.

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American Trypanosomiasis (AT) is an infectious parasitic disease produced by the protozoa Trypanosoma cruzi (T. cruzi). Infection is acquired by vectorial via but can also be transmitted congenitally, by ingestion of an infected host, by transfusion with contaminated blood or transplant of organs from an infected donor. Currently, AT is widely distributed from the South of the United States to South America. In Mexico, the presence of the parasite has been reported throughout the country where several reservoirs such as dogs, opossums, rats and cats have been identified. Yucatan is in the south-east of Mexico where AT is endemic and has been reported since 1940s. There is little information about the role of pigs as reservoirs of T. cruzi. The frequency of specific antibodies against T. cruzi was determined in fattening pigs from Yucatan, Mexico. After sampling in the 3 main areas of pig production in the state, IgG ELISA and Western blot were performed to identify seropositive cases. Association of farm size, farm area and production system with infected pigs was evaluated. From 273 sampled pigs, 5.4% (n = 15) positive cases were found. No association with evaluated factors and infected pigs was found. Pigs are also reservoirs of T. cruzi in the studied area. These findings are considered important to improve vectorial control in the area in order to avoid the parasite infection in animal populations destined for human consumption and avoid further transmission to humans.

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The objective of this study was to determine the seasonal ovarian activity of domestic queens under the tropical conditions of southern Mexico (19° 30' and 21° 35' N latitude). A total of 250 reproductive tracts were evaluated, and the frequencies of queens with ovarian activity (oestrus, dioestrus or pregnant), anoestrus, pregnancies and ovulation rate by season of the year (spring, summer, autumn and winter) were calculated. Ovarian activity was observed throughout the year, but a significant reduction occurred during the winter; conversely, anoestrous cases were lower during spring and showed an increase during summer, autumn and winter. Pregnancies were most common during spring and decreased during summer, autumn and winter. Ovulation rate varied from 2.5 to 2.8 during the four seasons. Cystic endometrial hyperplasia, pyometra and ovarian follicular cysts were the most common lesions found. The results demonstrate that domestic queens are not seasonal in tropical conditions, but a clear effect of the season of the year reducing ovarian activity is present.

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The presence of cats in the farms is considered a risk factor for the infection of pigs with Toxoplasma gondii (T. gondii). Cats eliminate oocysts that contaminate food, water and promote the infection of host reservoir such as rodents and birds among others that are also involved in the infection of pigs. The objective of this study was to assess the dynamic of infection of T. gondii in seronegative weaned pigs from weaning to 20 weeks of age from two farms from an endemic region, one with high and low density of cats. A cohort study was performed in 64 pigs, 31 newly weaned pigs on a farm with a high density of cats (FA) and 33 newly-weaned pigs on a farm with a low density of cats (FB). Blood samples were collected every 14 days to determine the presence of IgG antibodies against T. gondii in the serum using an indirect ELISA test. True incidence rate (TIV), cumulative incidence (AI) and relative risk (RR) was calculated. The age of seroconversion was determined by using survival tables; both farms were compared with Long-Rank test. In FA 97.5% of the pigs seroconverted at the second sampling and 100% at the third sampling, while in the FB all pigs seroconverted to the fourth sampling. The TIV was 0.67 and 0.43 for FA and FB respectively, during the first four weeks at risk. A RR of 1.5 (1.04-2.39) was obtained (p<0.05). Animals of the FA had a higher risk of infection compared with the FB, however, all animals included in the study had contact with the agent. Infection with T. gondii was rapidly distributed in both farms, regardless of the relative density of cats observed during the study. These results suggest a high environmental contamination with oocysts in the facilities of both farms probably due to the fact that T. gondii infection is endemic in the area where the farms are located, allow proper establishment of the etiological agent. The points of prevention and control strategies to avoid exposure of pigs to T. gondii in an endemic area should focus on the control of cats and rodents.

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American trypanosomiasis is an important zoonotic disease which affects more than 15 million persons in America. In Mexico, Chagas' disease is widely distributed in the country mostly in states with tropical weather conditions, including Yucatan. A cross-sectional study was performed on serum samples from 35 dogs and their owners (n = 75) from the south area of Merida city. Specific IgG antibodies against Trypanosoma cruzi (T. cruzi) using enzyme-linked immunosorbent assay, Immunofluorescence antibody test (IFI) and Western blot (WB) were detected. The overall percentage of seropositivity was 34% in dogs and 8% for sampled owners. Some owners brought samples of insect vectors found in their households, which were kept with BALB/c mice as a source of food and which were evaluated by polymerase chain reaction. All tested insects were positive to T. cruzi, and BALB/C mice were IFI and WB positive after 45 days in contact with these vectors. Further investigations showed that there is a high risk of infection with Chagas disease in dogs which spend the night outdoors (P < 0.05), with low body conditions score and older dogs. Risk factors identified to be associated to the infection in owners were gender and occupation (bricklayers, P < 0.05). The presence of seropositive dogs in houses where vectors are well-adapted represents a high risk for humans to become infected when bitten by a vector infected by a positive dog.

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A cross sectional survey was performed to identify gastrointestinal helminths and protozoans in naturally infected horses from the biosphere reserve known as "La Sierra Madre de Chiapas", Mexico (El Triunfo and La Sepultura). During a three-year survey, fecal samples from 90 horses and parasites from 2 necropsied animals were collected. Five families from the Nematoda class: Ascaridae, Kathlanidae, Oxyuridae, Strongylidae and Trichostrongylidae were found, whereas, only one family from the class Cestoda, was observed: Anoplocephalidae. One family from the class Insecta, was observed: Gasterophiilidae. The number of species of parasites ranged from 13 to 18 with an average of 15 per animal. Adult parasites were recovered from the large intestine luminal contents at necropsy. Species recovered included: Strongylus vulgaris, S. equinus, S. edentatus, Oxyuris equi, Parascaris equorum, Coronocyclus coronatum, C. labiatus, C. labratus, Cyathostomum tetracanthum, Cylicocyclus insigne, C. leptostomus, Cylicodontophorus bicoronatus, Cylicostephanus asymetricus, C. bidentatus, C. minutus, C. longibursatus, Petrovinema poculatum, Poteriostomum imparidentatum, Cylicostephanus goldi, Tridentoinfundibulum gobi, Triodontophorus serratus and T. tenuicollis. One species of Diptera were recovered from stomach and identified: Gasterophilus intestinalis. Furthermore, different species of protozoa were recovered from fresh horse-dung and identified in four classes: Sporozoa, Litostomatea, Ciliasida and Suctoria. Nine families: Cryptosporidiidae, Eimeriidae, Balantidiidae, Buetschliidae, Blepharocorythidae, Cycloposthiidae, Spirodiniididae, Ditoxidae, Acinetidae; and 31 ciliates species were recorded: Allantosoma dicorniger, A. intestinalis, Alloiozona trizona, Blepharosphaera intestinalis, Blepharoprosthium pireum, Blepharoconus benbrooki, Bundleia postciliata, Didesmis ovalis, D. quadrata, Sulcoarcus pellucidulus, Blepharocorys angusta, B. cardionucleata, B. curvigula, B. juvata, B. uncinata, B. valvata, Cycloposthium bipalmatum, C. edentatum, C. scutigerum, Charonina equi, Ditoxum funinucleum, Spirodinium equi, Tetratoxum unifasciculatum, Triadinium caudatum, T. galea, T. minimum and Tripalmaria dogieli. Other ciliate observed and recorded was Ochoterenaia appendiculata. This study describes the biodiversity and distribution of common and new helminths and protozoas found in the gastrointestinal tract from horses in the biosphere reserve "Sierra Madre de Chiapas" in Mexico.

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Papaya seeds (Carica papaya Linn) have been found to have a significant effect on sperm characteristics in some mammals, including humans, but no studies have investigated the effects on dogs. In the present study, a significant decrease in sperm concentration was observed in a group of dogs treated with extracts from washed papaya seeds, but no decrease was observed in the group of dogs treated with non-washed seeds. An important effect of extract components from washed seeds such as fatty acids is probably involved in the reduction of sperm production because of Sertoli cell damage, as has been suggested for langur monkeys. Dilution of the active components in the non-washed papaya seeds or interference with some of the components may reduce the expected effect on spermatogenesis. This first report on the effects of a chloroformic extract of papaya seeds in dogs suggests that an increased dose is necessary to achieve azoospermia.

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Canine ehrlichiosis is a disease produced by the rickettsial organism Ehrlichia canis. Reported prevalence may vary greatly depending on the test and sampling method used. For the serological detection of antibodies against E. canis, the indirect immunofluorescence antibody test (IFA) is considered the gold standard. However, other available serological techniques such as the indirect immunoperoxidase technique (IPT) have not yet been tested and may be efficient in detecting specific antibodies. Prevalence found (8.7% and 8.1% for IFA and IPT, respectively) was lower than previously reported in the studied area despite the well-established technique used for the determination of antigens. A kappa value of 0.958 (95% CI 0.9-1.0) was found with a sensitivity and specificity for IPT of 92.59% (95% CI 80.8-99.9) and 99.9% (95% CI 99.8 -100), respectively. The positive predictive value was 99.9% and the negative predictive value was 99.29%. The IPT technique can be used safely for serological determination of E. canis antibodies.

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The aim of this study was to determine the frequency and type of renal lesions associated with positive titers against Leptospira sp. in a stray dog population. Three hundred fifty pairs of kidneys and an equal number of serum samples were collected from dogs captured by the dog pound of Merida, Yucatan, Mexico. Euthanasia of dogs was performed following the regulations of the Official Mexican Health Ministry (NOM-033-ZOO-1995). Serum samples were evaluated with the microscopic agglutination test, and tissue samples were processed and fixed in paraffin. After staining with hematoxylin and eosin, the frequency of renal lesions was determined and classified. As an additional evaluation, samples with interstitial nephritis were stained by the Warthin-Starry method in order to observe the presence of spirochete forms that could be morphologically compatible with Leptospira spp. We found that 98% of cases presented at least one type of lesion. The main histological lesions found were mesangial proliferative glomerulonephritis (MPGN) in 63.7% (n= 223), mesangial proliferative glomerulonephritis and interstitial nephritis (MPGN+IN) in 34% (n= 119), nephrosclerosis in 0.57% (n= 2), mesangial glomerulonephritis in 0.28% (n= 1), and interstitial nephritis (IN) in 0.28% (n= 1). Thirty-four percent (n= 122) of the dogs were seropositive to Leptospira sp., mainly against serovar canicola. Among dogs with IN (alone or associated with MPGN) (n= 120), 49.1% were seropositive to Leptospira sp., but only 17% of them showed spirochete forms compatible with the bacteria. A statistical association between seropositive dogs and the presence of MPGN+IN was determined (P < 0.0001; odds ratio 2.7, confidence interval 1.7-4.5). We concluded that the frequency of renal lesions found in this study is high and L. canicola is probably the most common circulating serovar in dogs from this area. Dogs that have been in contact with Leptospira spp. have a higher risk of developing renal lesions of the type MPGN+IN.

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The objectives of this study were to determine the annual reproductive pattern and to estimate the frequency of reproductive pathologies in female mongrel stray dogs under tropical conditions. The genital tracts of 300 mongrel bitches from a municipal dog pound were examined post-mortem from January to December 2003. Season of the year, age, size, and body condition score (BCS) were recorded for each dog. The year was divided into three seasons: warm-dry (March-June), warm-humid (July-October), and fresh-humid (November-February). Distribution of estrus periods was not influenced by any of the factors studied (i.e. season, age, size, BCS). A significantly lower number of pregnancies were recorded during the warm-dry season, probably as a consequence of embryo resorption. Underweight animals had a significantly lower percentage of pregnancies than bitches of ideal BCS. More ovulations per bitch occurred during the warm-humid season than during the other seasons of the year, probably due to climatic factors. Bitches of medium and large size had more ovulations than those of small size. Of the 300 bitches examined, 43.5% had one or more genital pathologies. The most frequent pathologies found in the ovary, uterus, and vagina were epoöphoron cysts (6.7%), serosal inclusion cysts (5.0%), and transmissible venereal tumors (15.3%), but the capacity of the females to come into estrus or to become pregnant was not affected by these conditions. We concluded that stray domestic bitches in the tropics were not seasonal breeders, but their reproductive pattern was apparently modified by environmental factors such as temperature and probably photoperiod. Although several bitches in the present study had reproductive pathology, the most prevalent pathologies did not adversely reproductive capacity.

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The objective of this study was to estimate the prevalence of and risk factors for pathological conditions of the reproductive organs in stray dogs under tropical conditions. Three hundred and eighteen dogs were examined post-mortem in the period from 1 July 2002 to 30 June 2003. Before killing, a blood sample (from the cephalic vein) for testosterone assay was taken. Pathological conditions of the reproductive organs were found in 135 of the dogs (42.5%) and in 175 of the testes (64.8%). The most frequent pathologies found were testicular degeneration, cryptorchidism, testicular hypoplasia and testicular tumours (in 15.1%, 6.6%, 6.6% and 5.4% of the dogs and 15.1, 4.6, 6.0 and 3.5 of the testes, respectively). Transmissible venereal tumour (TVT) was seen in 5.4% of the dogs. Testicular degeneration was more common in old dogs and underweight dogs (p < 0.05). Testicular tumours were 14.3 times more common in cryptorchid dogs. Age was another important factor for the development of testicular tumours (p < 0.05). Lower levels of testosterone concentration (p < 0.05) were observed in dogs with advanced testicular degeneration (0.7 +/- 0.8 nM), dogs with hypoplastic testicles (0.8 +/- 0.9 nM) and dogs with one degenerated and one retained testis or with bilateral cryptorchidism (1.2 +/- 0.9 nM) compared to dogs with one or two normal testes (7.0 +/- 5.5 nM). Testicular volume and weight were significantly lower in degenerated, hypoplastic and retained testes compared with the contralateral normal testis. Some spermatogenic activity was found in three of the retained testes, producing oligozoospermic smears with a high percentage of sperm abnormalities. No comparable epidemiological data about male pathological conditions of the reproductive organs in the dog is available. The prevalence found in this study, yet, appears high.

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Two studies were performed to determine annual reproductive patterns in stray male dogs in the tropics. In Study 1, four dogs housed individually outdoors were monitored once monthly for 12 months, including collection and assessment of semen, measurements of scrotal width, and determination of serum testosterone and prolactin concentrations. In Study 2 (conducted concurrently), a single blood sample (for serum testosterone concentration) was collected from 220 clinically healthy dogs, and after euthanasia, scrotal width and morphology of epididymal sperm were determined. The year was divided into three seasons: warm-dry (March to June); warm-humid (July to October) and fresh-humid (November to February). In Study 1, scrotal width, ejaculate volume, sperm count and motility were significantly lower during the fresh-humid season and sperm midpiece abnormalities were significantly more common during the warm-humid and fresh-humid seasons. Serum testosterone concentrations remained constant during the year. Prolactin concentrations did not differ significantly among seasons, but had a well-defined increase from the beginning of March to the end of August. In Study 2, sperm morphology was similar to in Study 1 and serum testosterone concentrations varied nonsignificantly during the year. Environmental factors, e.g. daylength may have influenced circannual changes in prolactin secretion. Seasonal variations in some reproductive tract and seminal traits were significant but of small magnitude and the percentage of morphologically normal sperm did not vary significantly among seasons. In conclusion, healthy male dogs constantly produced sperm and were apparently fertile throughout the year.

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OBJECTIVES: To determine the prevalence of fetal resorption in 77 pregnant bitches and its association with the age, size and body condition score of the bitch and to determine the frequency of transuterine migration of embryos. METHODS: The number and position of embryos or fetuses and placental zones were registered postmortem. The prevalence of fetal losses was determined by comparing the number of corpora lutea with the number of embryos or fetuses in each bitch. The prevalence of transuterine migration of embryos was determined by correlating the number of fetuses in each horn and the number of corpora lutea in the ipsilateral ovary. RESULTS: Fetal resorption zones were demonstrated in 42.9 per cent of the bitches. However, pregnancy continued in 84.9 per cent of cases. Of the fetal losses, 25.9 per cent were determined by counting the number of corpora lutea and viable embryos or fetuses. Young bitches tended to have a higher probability of fetal resorption than adult bitches (P<0.06). The presence of fetal resorption zones was not associated with size or body condition. Transuterine migration of embryos had occurred in 15.8 per cent of the bitches. CLINICAL SIGNIFICANCE: This study demonstrated that embryo resorption is a normal event in the bitches, with a higher prevalence than previously thought.

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A Burdizzo castrator was evaluated for the neutering of dogs. Histological and morphological changes of spermatic cells and peripheral serum testosterone after challenge with a GnRH-analogue (gonadorelin) were assessed. There was a control group (G1), a surgically castrated group (G2) and a Burdizzo group (G3) divided in two, G3a receiving two crunches in each spermatic cord and G3b receiving one crunch in each spermatic cord. Sixteen days after application of the Burdizzo blood samples were taken from the dogs at 30 min interval during 2 h; after the second sample the dogs were treated with 1 mug/kg body weight of gonadorelin i.v. The same protocol of gonadorelin challenge was performed in G1 and G2 dogs. The G2 dogs were surgically castrated after the second blood sample, before the gonadorelin treatment, and the G1 dogs after the last blood sample. The excised gonads were examined histologically, and sperm smears were prepared from the caudae epididymidis. The testes and plexus pampiniformis of the G1 and G2 dogs had a normal histological appearance, and they had morphologically normal epididymal sperm cells. In all G3 dogs, there was an acute fibrosis with an inflammatory reaction in the plexus pampiniformis. The testes from the G3a dogs showed diffuse areas of infarction and degeneration of the parenchyma. Similar but less diffuse lesions were seen in group 3b dogs. The deferent ducts from all G3 dogs showed vasitis and/or sperm granulomas. Azoospermia or sperm malformations were observed in the epididymal smears from the G3 dogs. Testosterone concentration in the G1 dogs increased after gonadorelin application (p < 0.0001). The G2 dogs had basal testosterone levels after castration (p < 0.001) and did not respond to gonadorelin. Groups 3a and b showed a slight but non-significant increase in testosterone concentration after gonadorelin challenge, supposedly due to the reduction of testicular blood flow and loss of testicular interstitial tissue.

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