RESUMO
Puppy and kitten survival over the first weeks is particularly dependent on colostrum, a specific secretion of the mammary gland produced during the first 2 days post-partum. Colostrum is a source of nutrients and immunoglobulins. It also contributes to the digestive tract maturation. Colostrum differentiates from milk mainly based on its concentration in immunoglobulins G: 20-30 g/L in dog colostrum, 40-50 g/L in cats' vs <1 g/L in milk. IgG concentration rapidly drops after parturition (-50% in 24 hr). Immune quality of colostrum is highly variable between bitches, with no relationship with maternal blood IgG level, dam's age, breed size or litter size. In addition to systemic immune protection, colostrum also plays a major role for local digestive protection, due to IgA, lysozyme, lactoferrin, white blood cells and various cytokines. Energetic concentration of canine and feline colostrum is not superior to that of mature milk. It depends on colostrum fat concentration and is affected by breed size (higher in breeds <10 kg adult body weight). As puppies and kittens are almost agammaglobulinemic at birth, transfer of IgG from their digestive tract into their bloodstream is crucial for their survival, IgG absorption ending at 12-16 hr after birth. Energetic supply over the two first days of life, as evidenced by growth rate over the two first days of life, also affects risk of neonatal mortality. Early and sufficient suckling of colostrum is thus the very first care to be provided to newborns for their later health and survival.
Assuntos
Gatos , Colostro/imunologia , Colostro/fisiologia , Cães , Fenômenos Fisiológicos da Nutrição Animal , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Colostro/química , Citocinas/análise , Ingestão de Energia , Feminino , Imunoglobulina A , Imunoglobulina G , Lactoferrina/análise , Muramidase/análise , Especificidade da EspécieRESUMO
The objective of this study was to describe efficiency of reproduction of purebred dogs in field breeding conditions, from mating to weaning in France. Data were collected between 2010 and 2014 in 5,667 French breeding kennels via a reproduction management software (Breeding Management System, Royal Canin, Aimargues, France). Effect of breed size (Mini: adult body weight <10 kg; Medium: 10-25 kg; Maxi: 25-40 kg; Giant: >40 kg), age of dam and male on pregnancy rate, abortion rate and litter size were evaluated by multivariable models. Data on 45,913 heats (all with mating), from 27,221 bitches from 248 breeds, were analysed. At mating, mean age (±SD) was 3.1 ± 1.8 years for bitches and 3.3 ± 2.0 for males. Males originated from the same kennel as the females in 88.5% of the matings. Based on breeder's evaluation of the pregnancy status, pregnancy rate (number of pregnant females based on breeders declaration/number of heats) was 87.8% and abortion rate was 6.8%. Finally, 81.9% of the mated females gave birth to a litter. On 37,946 litters (204,537 puppies), mean litter size was 5.4 ± 2.8 puppies (range 1-24), which was influenced by breed size and dam age (p < .0001). Stillbirth rate was 7.4% and puppy mortality rate (stillbirth + mortality until 2 months of age) was 13.4%. Prolificacy and puppy mortality rates were affected by breed size and within a breed size, by breed. Despite probable approximations (as data originate from breeders declaration), this large-scale analysis provides reference values on reproductive performance in dogs.
Assuntos
Cães/fisiologia , Reprodução/fisiologia , Aborto Animal/epidemiologia , Envelhecimento , Animais , Tamanho Corporal , Cruzamento , Doenças do Cão/epidemiologia , Doenças do Cão/mortalidade , Feminino , França , Tamanho da Ninhada de Vivíparos , Masculino , Análise Multivariada , Gravidez , Valores de Referência , Estações do Ano , Especificidade da Espécie , Natimorto/epidemiologia , Natimorto/veterinária , DesmameRESUMO
Reproduction management and performances are evaluated in the feline species only through a limited number of animals and studies. Our objective was to provide reference figures in purebred cats, from a large-scale sample. Data were collected from an online software dedicated to cattery management (Breeding Management System®, BMS, Royal Canin, Aimargues, France). Information was recorded on a voluntary basis by French breeders between 2011 and 2014. Data were anonymously transferred for analysis. A total of 9,063 oestrous periods (in contact with a male) from 5,303 queens (45 breeds) were recorded from 1,521 breeders. Most matings (70.1%) occurred during increasing day length periods. The mean age at mating (±SD) was 2.7 ± 1.6 years for queens and 2.9 ± 1.9 years for tomcats. Pregnancy rate (based on breeders declaration) was 85.2%. Among queens declared pregnant, 8.4% failed to maintain pregnancy. Globally, 78% of the mated females gave birth to 28,065 kittens within 7,075 L. Mean litter size was 4.0 ± 1.9 kittens among which 8.5% were stillborn. Neonatal and paediatric mortality rate was 8.2%. In total, 16.0% of kittens born died before weaning. The results of this study are based on the largest feline database ever analysed. The figures collected can thus be used as reference to define average reproductive performances in numerous breeds for cat breeders. Further analysis will identify factors influencing reproductive performances and early mortality in the feline species.
Assuntos
Gatos/fisiologia , Reprodução/fisiologia , Aborto Animal/epidemiologia , Animais , Animais Recém-Nascidos , Cruzamento , Doenças do Gato/epidemiologia , Doenças do Gato/mortalidade , Ciclo Estral , Feminino , França , Tamanho da Ninhada de Vivíparos , Masculino , Gravidez , Natimorto/veterinária , DesmameRESUMO
Colostrum and milk are complex mammary secretions providing the puppy with many nutritional and immunological factors, which play a crucial role for its correct development and survival. In the case of colostrum and/or milk intake deficiency, puppies are at increased risk of infectious diseases. This work reviews the various nutritional hyperimmune supplementations proposed to provide a passive immune protection and to positively impact puppies' health. Some strategies rely on canine immunoglobulins: canine colostrum banking and canine serum/plasma supplementation. Others involve heterologous sources of antibodies and other immune factors: bovine colostrum or hyperimmune egg powder. Among the different solutions evaluated from birth to weaning, canine plasma and hyperimmune egg powder showed promising beneficial effect on puppies' health. Canine plasma seems to positively impact not only growth (increased growth during the neonatal period), but also digestive health (higher species richness of intestinal microbiota) and the general health (tendency of lower morbidity). Puppies supplemented with hyperimmune egg powder presented increased neonatal growth and decreased risk of canine parvovirus infection. Nevertheless, natural canine maternal colostrum and milk ingestion remains the optimal guarantee for puppies' health and survival, as a source of immunity, energy and growth factors.
Assuntos
Animais Recém-Nascidos/fisiologia , Cães/fisiologia , Nível de Saúde , Imunidade , Fenômenos Fisiológicos da Nutrição Animal , Animais , Animais Recém-Nascidos/imunologia , Bovinos , Colostro , Suplementos Nutricionais , Doenças do Cão/microbiologia , Doenças do Cão/prevenção & controle , Cães/imunologia , Feminino , Imunoglobulinas , Infecções/veterinária , Leite , SoroRESUMO
BACKGROUND: In retrospective case-control studies performed following nuclear tests or nuclear accidents, individual thyroid radiation dose reconstructions are based on fallout and meteorological data from the residential area, demographic characteristics, and lifestyle as well as dietary information. Collecting the latter is a controversial step, as dietary declarations may be affected by the subjects' beliefs about their risk behavior. This report analyses the potential for such bias in a case-control study performed in eastern France. METHODS: The study included 765 cases of differentiated thyroid carcinoma matched with 831 controls. Risk perceptions and beliefs of cases and controls were compared using Chi2 tests and differences in dietary reports were analyzed using a two-way ANOVA. RESULTS: In general, atmospheric pollution and living near a nuclear power plant were the two major risks that may influence thyroid cancer occurrence cited by cases and controls. When focusing in particular on the consequences of the Chernobyl accident, cases were more likely to think that the consequences were responsible for thyroid cancer occurrence than controls. Vegetable consumption during the two months after the Chernobyl accident was correlated with the status of subjects, but not to their beliefs. Conversely, consumption of fresh dairy products was not correlated with the status or beliefs of subjects. CONCLUSION: We found no evidence of systematic bias in dietary reports according to the status or beliefs held by subjects about the link between thyroid cancer occurrence and Chernobyl fallout. As such, these dietary reports may be used in further studies involving individual dosimetric reconstructions.
Assuntos
Acidente Nuclear de Chernobyl , Registros de Dieta , Comportamento Alimentar/psicologia , Contaminação Radioativa de Alimentos , Percepção , Cinza Radioativa , Neoplasias da Glândula Tireoide/epidemiologia , Adolescente , Adulto , Viés , Estudos de Casos e Controles , Criança , Desastres , Feminino , França/epidemiologia , Humanos , Masculino , Centrais Nucleares , Inquéritos Nutricionais , Cinza Radioativa/análise , Cinza Radioativa/estatística & dados numéricos , Estudos Retrospectivos , Fatores de Risco , Comportamento de Redução do Risco , Adulto JovemRESUMO
Luteolysis before the time of maternal recognition of pregnancy is one cause of low fertility in high-producing dairy cows. The objective of this study was to assess whether induction of a secondary corpus luteum (CL) late in the luteal phase would delay the time of luteolysis. Twenty high-producing Holstein cows were synchronized to ovulation (Day 0) with the Ovsynch protocol and received hCG (1500 IU im) on Day 12. Corpora lutea formation (as evaluated by ultrasonography) and plasma P4 concentrations were monitored from Days 4 to 36. hCG treatment induced the formation of one secondary CL (CL2) in 11 of 20 cows (55%) from the dominant follicle (mean diameter: 14.2 ± 0.9 mm) of two-wave (3/11) and three-wave (8/11) cycles. The maximal diameter of the CL2 (23.3 ± 1.9 mm) was reached approximately 6 days after hCG treatment and was correlated with its structural lifespan (p < 0.01). Cows that formed a CL2 after hCG had higher mean plasma P4 concentrations on Day 14 (+4.5 ng/ml) and Day 18 (+3.0 ng/ml) compared with cows without CL2 (p < 0.05). The structural regression of CL2 begun approximately 8 days after that of the CL1, and the median time at which the first drop in circulating P4 levels occurred was later in cows that formed a CL2 than in those that did not (Day 26 vs Day 18; p < 0.01). Thus, the induction of a CL2 by hCG on Day 12 might reduce the risk of premature luteolysis in high-producing dairy cows after insemination.
Assuntos
Bovinos/fisiologia , Gonadotropina Coriônica/farmacologia , Corpo Lúteo/fisiologia , Luteólise/fisiologia , Ovulação/fisiologia , Animais , Busserrelina/administração & dosagem , Busserrelina/farmacologia , Cloprostenol/administração & dosagem , Cloprostenol/farmacologia , Ciclo Estral , Feminino , Lactação/fisiologia , Gravidez , Estações do AnoRESUMO
Important reproductive events take place in the canine oviduct in the presence of increasing concentrations of progesterone (P4). To investigate the potential effects of P4 on the canine oviduct, the expression of nuclear (PR) and membrane (PGRMC1 and 2, mPRα, ß and γ) P4 receptors was studied by quantitative RT-PCR and immunohistochemistry. Oviducts were collected from Beagle bitches after the onset of pro-oestrus and before the LH peak (Pre-LH), after the LH peak and before ovulation (Pre-ov) and on Days 1, 4 and 7 post-ovulation (n=6 bitches/stage). PR mRNA concentrations decreased from Pre-LH to Day 7 in the ampulla and isthmus, whereas both PGRMC1 and 2 mRNA levels increased over the same period. The main change in mPR expression was an increase in mPRß and γ mRNAs at Day 7 in the isthmus. Furthermore, PR proteins were expressed in the nuclei of luminal epithelial, stromal and muscular cells, whereas the expression of PGRMCs and mPRs was primarily cytoplasmic and localised in the luminal epithelium. The immunostaining for PR decreased at Day 4 in the stroma and muscle, whereas it remained strong in the epithelium from Pre-LH to Day 7. PGRMC1 staining was strong at Days 4 and 7 whereas PGRMC2 was highly expressed from Pre-ov to Day 7. The most intense immunostaining signals for all three mPRs were observed at Day 7. Our results strongly support the hypothesis that P4 is an important regulator of oviductal functions in the bitch through complementary classical and non-classical P4 pathways.
Assuntos
Cães , Tubas Uterinas/metabolismo , Tubas Uterinas/ultraestrutura , Expressão Gênica , Ovulação/metabolismo , Receptores de Progesterona/genética , Animais , Membrana Celular/química , Núcleo Celular/química , Estradiol/sangue , Tubas Uterinas/química , Feminino , Naftoquinonas , Progesterona/sangue , Progesterona/fisiologia , RNA Mensageiro/análise , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Receptores de Progesterona/análiseRESUMO
In contrast to endometritis, now diagnosed by cytological examination, the effect of endocervical inflammation on reproductive performance has been inadequately investigated. In this study, endocervical and endometrial cytological specimens were collected from 168 Holstein cows between 21 and 60 days in milk (DIM) to investigate the prevalence of endocervical inflammation and effect on days to conception. Statistical analyses were stratified based on DIM at examination (<35 vs. ≥35 DIM). Endocervical inflammation with ≥5% neutrophils before 35 DIM (disregarding the level of endometrial inflammation) was associated with decreased hazard of pregnancy within 300 DIM (adjusted hazard ratio, 0.4; 95% confidence interval: 0.3-0.8). A decrease in hazard of pregnancy was observed when >6% neutrophils were counted in endometrial smears (adjusted hazard ratio, 0.4; 95% confidence interval: 0.2-0.7). The study suggested an additive effect of combined endocervical and endometrial inflammation on the hazard of pregnancy within 300 DIM. Using the thresholds of 5% neutrophils for the cervix and 6% neutrophils for the uterus, 11% of the cows examined before 35 DIM presented cervicitis only, 13% were affected by endometritis only, and 32% suffered from both endometrial and endocervical inflammation. The presence (absence) of cervicitis was not indicative of the presence (absence) of endometritis. This study showed that in addition to uterine inflammation, endocervical inflammation in early lactation affects conception. Thus, the global evaluation of genital tract health may be more beneficial for reproductive performance than that of endometrial inflammation.
Assuntos
Doenças dos Bovinos/fisiopatologia , Fertilização , Cervicite Uterina/veterinária , Animais , Bovinos , Doenças dos Bovinos/patologia , Endometrite/complicações , Endometrite/veterinária , Feminino , Lactação , Neutrófilos/patologia , Fatores de Tempo , Cervicite Uterina/complicações , Cervicite Uterina/diagnósticoRESUMO
Heat detection is a key factor in the profitability of dairy herds. However, this detection demands a significant part of the breeder's working time and is made difficult by the short duration and the discrete behavioural changes associated with oestrus in modern dairy cows. Progress has been made in monitoring cow with electronics, biosensors and computer. As a result, automated heat detection systems have been developed. Currently available tools are automated detectors of standing heat, activity-metres and automated in-line systems measuring milk progesterone. Camera-software systems and monitoring of body temperature are being developed and may also be used as heat detection tools. The heat detection rate of most systems is above 80% with a specificity of detection generally higher than 90%. The accuracy, however, may vary considerably depending on the tool and model developed. The initial investment of several thousands of euros required for these automated systems becomes a source of profit in large herds, provided the recorded data are properly managed.
Assuntos
Automação , Bovinos/fisiologia , Detecção do Estro/instrumentação , Detecção do Estro/métodos , Estro/fisiologia , Animais , FemininoRESUMO
In the bitch, oocyte maturation, sperm storage, fertilization and early embryo development take place within the oviducts under high and increasing circulating progesterone concentrations. To investigate the potential effects of progesterone on the canine oviduct, nuclear progesterone receptors (PR) were localized. Oviducts were collected by ovariectomy from adult Beagle bitches during anestrus, after the onset of proestrus but prior to the Luteinizing Hormone (LH) peak (Pre-LH), after the LH peak but before ovulation (Pre-ov) and on Days 1, 4 and 7 after ovulation (n = 3 bitches per stage). The cellular distribution of PR was studied by immunohistochemistry (IHC) in the ampulla, isthmus and tubal part of the utero-tubal junction (UTJ). Plasma progesterone and 17ß-oestradiol were assayed on the day of surgery. PR were specifically expressed in the nuclei of epithelial, stromal and muscular cells in the ampulla, isthmus and UTJ. The IHC scores did not vary from one oviductal region to another. However, the epithelium displayed higher scores than the stroma at anestrus, Pre-ov, Days 4 and 7, and also higher scores than muscle at Days 4 and 7 (p < 0.05). Immunohistochemistry scores in the stroma and muscle decreased at Days 4 and 7 compared with previous stages (p < 0.05). Furthermore, muscular IHC scores were positively correlated with circulating 17ß-oestradiol concentrations and negatively correlated with circulating progesterone concentrations (p < 0.05). In conclusion, PR were identified in the canine oviduct, with differences in expression between tissues and times around ovulation, suggesting that progesterone may regulate tubal functions and reproductive events in this species.
Assuntos
Cães/fisiologia , Imuno-Histoquímica/veterinária , Oviductos/fisiologia , Ovulação/fisiologia , Receptores de Progesterona/fisiologia , Animais , FemininoRESUMO
In canine species, in vitro maturation (IVM) rates of oocytes collected from anoestrous ovaries are low (<20%). Several IVM media have been tested without significant improvements. A critical step in the evaluation of culture conditions is the observation of the meiotic stage reached by the oocytes. The present study was designed to investigate the chromatin patterns of in vitro matured oocytes by visualizing Germinal Vesicle (GV) and Germinal Vesicle Breakdown (GVBD) structures at 72 h of IVM. Nuclear stages of 1678 oocytes were evaluated by confocal microscopy after IVM. 1204 oocytes were non-degenerated, and 94.4% were still immature and at GV stage. Five different patterns of chromatin configuration were observed. Higher percentages of oocytes with unmodified GV and with diffuse (58%; Type A) and filamentous chromatin (19%; Type B) were observed in comparison with those with modifications in the GV such as patched chromatin (12.5%; Type C), surrounded-nucleolus (3%; Type D) and in vivo type chromatin/fully grouped chromatin (2.5%; Type E). These results indicate that GVBD (absence of nucleolus, nucleus breakdown) is rarely observed in vitro. The percentage of type C-D-E GVs and MI (meiotic resumption) and of MII (completion of meiosis) can be used to evaluate meiotic resumption after IVM. Our results indicate that although a low number of in vitro matured oocytes exhibit the chromatin configurations observed in in vivo collected oocytes, chromatin changes in the GV can be induced during IVM.
Assuntos
Cromatina/ultraestrutura , Cães/fisiologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/fisiologia , Oócitos/ultraestrutura , Animais , Sobrevivência Celular , Células Cultivadas , FemininoRESUMO
In the bitch, oocyte meiosis resumption takes place in the oviduct. Using oocytes from anestrous bitches, in vitro maturation (IVM) generally gives very poor results. To investigate the contribution of oocyte competence to the low IVM yield, we compared in vivo maturation in an optimal environment with conventional IVM. A total of 418 grade 1 cumulus-oocyte complexes (COCs) from 10 anestrous bitches were transferred into the oviducts of recipient bitches either on Day -1 (n = 3 recipients), Day 0 (n = 2) or on Day +1 (n = 2) relative to ovulation. For each donor bitch, 20 grade 1 COCs were also cultured in vitro. After 72 h of in vivo or IVM, the nuclear stage of oocytes was determined after DNA and tubulin staining. Of the 154 oocytes recovered and examined after intratubal transfer, only 2% reached the metaphase I or II stage and 38.3% were degenerated. Oocytes cultured in vitro displayed a higher metaphase rate (7.6%, n = 170) and lower degeneration rate (12.9%) compared with transferred oocytes (p < 0.001). These results clearly demonstrate that the oocyte competence is the major limiting factor of IVM efficiency in the dog. Mimicking the tubal environment may thus not be sufficient to increase IVM yield in this species.
Assuntos
Anestro/fisiologia , Cães/fisiologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Meiose/fisiologia , Oócitos/fisiologia , Animais , Células Cultivadas , FemininoRESUMO
As puppies are born with very low immunoglobulin concentrations, they rely on passive immune transfer from ingested colostrum to acquire a protective immunity during the first few weeks of life. The purpose of this study was to describe the timing of gut closure in canine neonates. Twenty-two Beagle puppies received 3 ml of standardized canine colostrum at 0, 4, 8, 12, 16 or 24 h after birth using a feeding tube. Blood immunoglobulins G (IgG, M and A) were assayed 0, 4 and 48 h after colostrum ingestion. IgG absorption rate was significantly affected by the time of colostrum administration, and the IgG concentrations in puppies serum 48 h after administration were significantly higher when colostrum was ingested at 0-4 h of age than at 8-12 h or 16-24 h (1.68 ± 0.4, 0.79 ± 0.07 and 0.35 ± 0.08 g/l, respectively; p < 0.001). In the canine species, gut closure seems thus to begin as early as 4-8 h after birth and to be complete at 16-24 h. Consequently, this phenomenon appears to occur earlier in puppies than in most other species.
Assuntos
Animais Recém-Nascidos , Cães/crescimento & desenvolvimento , Absorção Intestinal/fisiologia , Intestinos/crescimento & desenvolvimento , Intestinos/fisiologia , Animais , Colostro , Imunoglobulina A/metabolismo , Imunoglobulina G/metabolismo , Imunoglobulina M/metabolismo , Fatores de TempoRESUMO
Reproductive physiology in dogs is quite unusual compared with that in other mammalian species. The peculiarities include the presence of numerous polyoocyte follicles, the ovulation of an immature oocyte (GV stage, non-fertilizable) and a peri-ovulatory period during which concentrations of circulating progesterone are particularly high. The aim of this review is to examine the unusual aspects of the reproductive physiology of dogs and how this relates to the clinical biology of this species.
Assuntos
Cães/embriologia , Cães/fisiologia , Embrião de Mamíferos/fisiologia , Oócitos/fisiologia , Folículo Ovariano/fisiologia , Ovulação/fisiologia , Animais , FemininoRESUMO
OBJECTIVE: Cerebral exteriorization through the cribriform plate during routine endotracheal suctioning (ETS) in a coughing patient without sedation with multiple complex skull base fracture; this case has never been reported in the literature. CLINICAL PRESENTATION: We report the case of a polytrauma patient admitted in our ICU with severe traumatic brain injury (TBI) and multiple complex skull base fractures. After 48 hours of neurocritical care and stable neuromonitoring parameters, sedation was stopped for neurological assessment. During this period and while routine ETS was being performed, brain herniation with exteriorization through the nose occurred with a concomitant ICP elevation. CONCLUSIONS: ETS can induce the coughing reflex and provoke a rise in ICP. It is a simple routine procedure that should be performed with great precautions in order to avoid iatrogenic complications, particularly in patients with skull base fractures, such as brain herniation as described in our patient.
Assuntos
Lesões Encefálicas Traumáticas/cirurgia , Base do Crânio/cirurgia , Fraturas Cranianas/cirurgia , Sucção/efeitos adversos , Idoso , Encéfalo/diagnóstico por imagem , Tosse/complicações , Tosse/etiologia , Evolução Fatal , Hérnia/etiologia , Humanos , Pressão Intracraniana , Imageamento por Ressonância Magnética , Masculino , Nariz , Base do Crânio/lesões , Tomografia Computadorizada por Raios X , TraqueiaRESUMO
In the field of molecular and epidemiological parasitology, characterization of fast evolving genetic markers appears as an important challenge to consider the diversity and genetic structure of parasites. The study of respective populations can help us to understand their adaptive strategies to survive and perpetuate the species within different host populations, all trying to resist infection. In the past, the relative monomorphic features of Echinococcus multilocularis, the causative agent of alveolar echinococcosis and a severe human parasitic disease, did not stimulate studies dealing with the genetic variability of Echinococcus species or respective populations. A recently developed, characterized and validated original multilocus microsatellite, named EmsB, tandemly repeated in the genome, offered an additional opportunity for this line of investigation. We have compiled in this review new insights brought by this molecular tracker on the transmission activity of Echinococcus among different hosts and at different geographical scales.
Assuntos
Equinococose/epidemiologia , Equinococose/parasitologia , Echinococcus/genética , Genômica , Repetições de Microssatélites , Animais , Echinococcus/classificação , Regulação da Expressão Gênica/fisiologia , HumanosRESUMO
Cystic echinococcosis (CE) is a widespread and severe zoonotic disease caused by infection with the larval stage of the eucestode Echinococcus granulosus sensu lato. The polymorphism exhibited by nuclear and mitochondrial markers conventionally used for the genotyping of different parasite species and strains does not reach the level necessary for the identification of genetic variants linked to restricted geographical areas. EmsB is a tandemly repeated multilocus microsatellite that proved its usefulness for the study of genetic polymorphisms within the species E. multilocularis, the causative agent of alveolar echinococcosis. In the present study, EmsB was used to characterize E. granulosus sensu lato samples collected from different host species (sheep, cattle, dromedaries, dogs, and human patients) originating from six different countries (Algeria, Mauritania, Romania, Serbia, Brazil, and the People's Republic of China). The conventional mitochondrial cox1 and nad1 markers identified genotypes G1, G3, G5, G6, and G7, which are clustered into three groups corresponding to the species E. granulosus sensu stricto, E. ortleppi, and E. canadensis. With the same samples, EmsB provided a higher degree of genetic discrimination and identified variations that correlated with the relatively small-scale geographic origins of the samples. In addition, one of the Brazilian single hydatid cysts presented a hybrid genotypic profile that suggested genetic exchanges between E. granulosus sensu stricto and E. ortleppi. In summary, the EmsB microsatellite exhibits an interesting potential for the elaboration of a detailed map of the distribution of genetic variants and therefore for the determination and tracking of the source of CE.
Assuntos
DNA de Helmintos/genética , Equinococose/parasitologia , Equinococose/veterinária , Echinococcus granulosus/classificação , Echinococcus granulosus/genética , Variação Genética , Repetições de Microssatélites , Argélia , Animais , Brasil , Camelus , Bovinos , China , Análise por Conglomerados , Cães , Echinococcus granulosus/isolamento & purificação , Genótipo , Humanos , Mauritânia , Romênia , Sensibilidade e Especificidade , Sérvia , OvinosRESUMO
In vitro maturation (IVM) of bitch oocytes is, to date, a very inefficient process, with common metaphase rates approximately 0-20% and mean degeneration rates approximately 20-30%. In other mammals, meiotic resumption is controlled in the cumulus-oocyte complex by the disappearance of the coupling between granulosa cells and the oocyte. The first aim of this study was to evaluate the influence on meiotic resumption of a mechanical denudation of the oocytes before maturation. The nuclear stage was determined by DNA staining with ethidium-homodimer-2 under confocal microscopy. Denuded (n = 318) and control (n = 378; no mechanical denudation) oocytes had similar degeneration rates (respectively 32.1 vs 28.6%). However, meiosis resumption rates were significantly higher for denuded oocytes (DO; 26.9 vs 17.8%). Secondly, we aimed to evaluate oocytes experiencing spontaneous denudation during the 72 h IVM period. Denuded oocytes, having lost cumulus cells on at least 75% of their perimeter (n = 440), were compared with surrounded oocytes (SO), with 100% of their perimeter surrounded by granulosa cells (n = 860). As above, the nuclear stage was determined by confocal microscopy, but cytoplasmic maturation was also evaluated through transmission electron microscopy. Degeneration rates but also meiosis resumption and metaphase rates were significantly higher for denuded than for SO (9.6 vs 2.8% for metaphase rate). Nevertheless, ultrastructurally, metaphase DO have scarcer organelles unevenly distributed, with smooth endoplasmic reticulum concentrated in aggregates in the cortical zone. Denudation, whether mechanical or spontaneous, is thus an inefficient mean to obtain metaphase II oocytes suitable for in vitro fertilization.
Assuntos
Células do Cúmulo/fisiologia , Cães , Oócitos/fisiologia , Animais , Núcleo Celular , Células Cultivadas , Citoplasma , Feminino , Meiose/fisiologiaRESUMO
This study was designed to describe, both quantitatively (morphometry) and qualitatively (histological differentiation), follicle and oocyte growth in the feline ovary. The ovaries of 43 cats were collected and processed for histology. The diameters of 832 follicle/oocyte pairs were measured, with and without zona pellucida (ZP), and a special emphasis was placed on the study of early folliculogenesis. Primordial, primary, secondary, pre-antral and early antral follicles were measured at 44.3, 86.2, 126.0, 155.6 and 223.8 microm in diameter respectively. A biphasic pattern of follicle and oocyte growth was observed. Before antrum formation, follicle (x) and oocyte (y) size were positively and linearly correlated (y = 0.500x + 20.01, r(2) = 0.89). Antrum formation occurred when the follicle reached 160-200 microm in diameter (when oocyte was at 102 microm). After antrum formation, a decoupling was observed, a minimal increase in oocyte size contrasting with a significant follicle development (y = 0.001x + 114.39, r(2) = 0.01). The pre-ovulatory follicle diameter was approximately 3500 microm and the maximal oocyte diameter was 115 microm. The ZP, absent in primordial and primary follicles, appeared at the secondary stage and reached almost 6 microm at the pre-ovulatory stage. These results suggest that (i) in feline ovary, follicle and oocyte growth pattern is similar to that observed in other mammals; (ii) the antrum forms in 160-200 microm follicles, which represents 5% of the pre-ovulatory diameter and (iii) the oocyte had achieved more than 90% of its maximal growth at the stage of antrum formation.
Assuntos
Gatos/anatomia & histologia , Gatos/fisiologia , Oócitos/fisiologia , Folículo Ovariano/fisiologia , Animais , Ciclo Estral , Feminino , Oócitos/citologia , Folículo Ovariano/anatomia & histologia , Zona Pelúcida/ultraestruturaRESUMO
PURPOSE: The delineation of volumes of interest can be a source of significant interobserver variability. The purpose of this study was to improve the homogeneity of delineation between oncologist-radiotherapists in the territorial departments of Nord and Pas-de-Calais (France) through discussions of clinical cases and the adoption of common published reference documents. MATERIALS AND METHODS: All eleven radiotherapy centres in the Nord and Pas-de-Calais departments of France participated. The localizations assessed to date included prostate, head and neck, breast and brain cancers. For each localization, the junior or senior physician(s) in charge of pathology delineated the volumes of interest according to their usual practices. Validated indices, including the Dice similarity coefficient, were used to quantify the delineation differences. The anonymized results were presented at two to three annual meetings. A second delineation of the clinical cases was then carried out to quantify homogenization. An evaluation of dosimetry practices was also conducted for prostate cancer. Wilcoxon assay matched data were used. RESULTS: Our work showed either satisfactory delineation concordance after the initial assessment or improved delineation concordance. For prostate cancer, the Dice similarity coefficient values were greater than 0.6 initially in two of the three clinical cases. For head and neck cancers, a statistically significant improvement was observed for only one of the clinical target volumes. More than half of the Dice similarity coefficient values were greater than 0.6 in the first comparison. The study of clinical cases of breast cancer allowed a homogenization of the delineation of five of the six lymph node clinical target volumes. The dosimetry study of prostate cancer allowed for a homogenization of practices. CONCLUSION: This work makes it possible to harmonize the delineation practices around validated standards. An extension to the entire Hauts-de-France region is planned.