Combined approach for characterization and quality assessment of rabbit bone marrow-derived mesenchymal stem cells intended for gene banking.

Rabbit mesenchymal stem cells (rMSCs) are promising agents for the preservation of genetic biodiversity in domestic rabbit breeds. However, rMSCs must meet certain requirements to be used for cryopreservation in animal gene banks. Currently, there are numerous discrepancies in the published data regarding the rMSC phenotype, which may complicate efforts to evaluate their purity and suitability for reuse after cryopreservation in gene and tissue banks. We propose a combined approach (flow cytometry, PCR, differentiation and ultrastructure studies) for the characterization and recovery of rMSCs after cryopreservation. Flow cytometric analyses of rMSCs confirmed the expression of CD29, CD44, vimentin, desmin and α-SMA. RT-PCR revealed the expression of other markers at the mRNA level (SSEA-4, CD73, CD90, CD105, CD146 and CD166). rMSCs showed efficient multilineage differentiation into adipo-, chondro- and osteogenic lineages, SOX2 expression (pluripotency) and typical MSC morphology and ultrastructure. The confirmed rMSCs were subsequently used for cryopreservation. Efficient recovery of rMSCs after cryogenic freezing was demonstrated by high cell viability, normal ultrastructure of reseeded rMSCs, high expression of CD29 and CD44 and lineage differentiation capacity. The proposed combined approach could be used for characterization, cryopreservation and recovery of rMSCs as genetic resources for native rabbit breeds.

Identification of Long Non-Coding RNAs Related to Skeletal Muscle Development in Two Rabbit Breeds with Different Growth Rate.

Skeletal muscle development plays an important role in muscle quality and yield, which decides the economic value of livestock. Long non-coding RNAs (lncRNAs) have been reported to be associated with skeletal muscle development. However, little is revealed about the function of lncRNAs in rabbits' muscle development. LncRNAs and mRNAs in two rabbit breeds (ZIKA rabbits (ZKR) and Qixin rabbits (QXR)) with different growth rates at three developmental stages (0 day, 35 days, and 84 days after birth) were researched by transcriptome sequencing. Differentially expressed lncRNAs and mRNAs were identified for two rabbit breeds at the same stages by DESeq package. Co-expression correlation analysis of differentially expressed lncRNAs and mRNAs were performed to construct lncRNA⁻mRNA pairs. To explore the function of lncRNAs, Gene Ontology (GO) analysis of co-expression mRNAs in lncRNA⁻mRNA pairs were performed. In three comparisons, there were 128, 109, and 115 differentially expressed lncRNAs, respectively. LncRNAs TCONS_00013557 and XR_518424.2 differentially expressed in the two rabbit breeds might play important roles in skeletal muscle development, for their co-expressed mRNAs were significantly enriched in skeletal muscle development related GO terms. This study provides potentially functional lncRNAs in skeletal muscle development of two rabbit breeds and might be beneficial to the production of rabbits.

Large-scale assessment of myxomatosis prevalence in European wild rabbits (Oryctolagus cuniculus) 60years after first outbreak in Spain.

Myxomatosis is a viral disease that affects European rabbits (Oryctolagus cuniculus) worldwide. In Spain, populations of wild rabbits drastically decreased in the 1950s after the first outbreak of myxomatosis. Since that first appearance, it seems to be an annual epizootic in Spain with periodic outbreaks, predominantly in summer and autumn. Taking into account rabbit population structure, abundance, and genetic lineage, this paper attempts to make a large-scale characterization of myxomatosis seroprevalence based on the immune status of 29 rabbit populations distributed throughout Spain, where O. cuniculus cuniculus and O. c. algirus, the two known rabbit subspecies, naturally inhabit. A total of 654 samples were collected between 2003 and 2009, and seroprevalence of antibodies against Myxoma virus (MYXV) was determined. Overall, our results revealed that 53% of the rabbit samples were positive to antibodies against MYXV. Newborn and juvenile rabbits were the most susceptible animals to the virus, with 19% and 16% seropositivity for newborn and juveniles, respectively, while adult rabbits were the most protected, with 65% of seropositive samples. This suggests that prevalence is negatively related to the proportion of newborn and juvenile rabbits in a population. Our results also showed that seroprevalence against MYXV tended to be higher in high-abundance populations. In contrast, no differences were detected in seroprevalence between rabbit subspecies. This study confirms that >60years since first outbreak, myxomatosis is an endemic disease in Spain. Based on the results, the establishment of a myxomatosis surveillance protocol is proposed.

Measurement of Tear Production in English Angora and Dutch Rabbits.

The purpose of this study was to establish normal values for tear production tests in different breeds of domestic rabbits. Healthy adult rabbits (n = 60; 120 eyes) of 2 different breeds (English angora and Dutch; n = 15 of each sex and breed) were used in this study. Tear production was measured by using the 1-min Schirmer tear test (STT), phenol red thread test (PRTT), and endodontic absorbent paper point tear test (EAPTT). In addition, horizontal palpebral fissure length was evaluated as a measure of ocular adnexal dimensions. Tear production (mean ± 1 SD) in English angora rabbits was 5.4 ± 1.6 mm/min according to the STT, 25.0 ± 2.7 mm in 15 s for the PRTT, and 18.8 ± 2.1 mm/min by the EAPTT; in Dutch rabbits, these values were 4.6 ± 1.2 mm/min, 23.6 ± 2.3 mm in 15 s, and 16.9 ± 1.7 mm/min, respectively. Only the EAPTT revealed a significant difference in tear production between English Angora and Dutch rabbits. These results provide reference values for tear production in English Angora and Dutch rabbits according to 3 different quantitative tear film assessment methods.

Growth traits of four maternal lines of rabbits founded on different criteria: comparisons at foundation and at last periods after selection.

The objective was to compare growth traits in four maternal lines of rabbits (A, V, H and LP), with the aim of understanding the consequence of the different foundation and selection processes on the growth performance of the lines. The lines are currently in the 43th, 38th, 22th and 8th generations, respectively. Two comparisons were performed. One compared the values of the lines at their foundation, using the complete data set, the full pedigree and a two-trait analysis, including data on the selection criteria, litter size. The other comparisons were done during the last period when all the lines were housed together with the same feeding and management. The numbers of records were 323 208 for weaning weight, and 300 553 for slaughter weight and average daily Gain (from 46 708 l). The pedigree file included 346 638 animals. The second analysis used only the data corresponding to each period, and the analysis was conducted using a one-trait model. The model was the same as that defined for the comparisons at the foundation, but the additive effects were excluded. The H and LP lines showed highest values for all the traits compared. In the last periods, a good agreement was observed between the estimated differences, computed with the complete model and data set, or computed with an incomplete model and only data from the comparison period. At last periods, the differences were smaller than at foundation. The importance of the correlated response in growth after selection for litter size at weaning or the importance of a non-programmed intramating selection for the growth traits can explain the changes since foundation.

A Century of Shope Papillomavirus in Museum Rabbit Specimens.

Sylvilagus floridanus Papillomavirus (SfPV) causes growth of large horn-like tumors on rabbits. SfPV was described in cottontail rabbits (probably Sylvilagus floridanus) from Kansas and Iowa by Richard Shope in 1933, and detected in S. audubonii in 2011. It is known almost exclusively from the US Midwest. We explored the University of Kansas Natural History Museum for historical museum specimens infected with SfPV, using molecular techniques, to assess if additional wild species host SfPV, and whether SfPV occurs throughout the host range, or just in the Midwest. Secondary aims were to detect distinct strains, and evidence for strain spatio-temporal specificity. We found 20 of 1395 rabbits in the KU collection SfPV symptomatic. Three of 17 lagomorph species (S. nuttallii, and the two known hosts) were symptomatic, while Brachylagus, Lepus and eight additional Sylvilagus species were not. 13 symptomatic individuals were positive by molecular testing, including the first S. nuttallii detection. Prevalence of symptomatic individuals was significantly higher in Sylvilagus (1.8%) than Lepus. Half of these specimens came from Kansas, though new molecular detections were obtained from Jalisco-Mexico's first-and Nebraska, Nevada, New Mexico, and Texas, USA. We document the oldest lab-confirmed case (Kansas, 1915), pre-dating Shope's first case. SfPV amplification was possible from 63.2% of symptomatic museum specimens. Using multiple methodologies, rolling circle amplification and, multiple isothermal displacement amplification in addition to PCR, greatly improved detection rates. Short sequences were obtained from six individuals for two genes. L1 gene sequences were identical to all previously detected sequences; E7 gene sequences, were more variable, yielding five distinct SfPV1 strains that differing by less than 2% from strains circulating in the Midwest and Mexico, between 1915 and 2005. Our results do not clarify whether strains are host species specific, though they are consistent with SfPV specificity to genus Sylvilagus.

A comprehensive linkage map and QTL map for carcass traits in a cross between Giant Grey and New Zealand White rabbits.

BACKGROUND: Genomic resources for the rabbit are still limited compared to many other livestock species. The genomic sequence as well as linkage maps have gaps that hamper their use in rabbit genome research. Therefore, the aims of this study were the improvement of existing linkage maps and the mapping of quantitative trait loci (QTL) for carcass and meat quality traits. The study was performed in a F2 population of an initial cross between Giant Grey (GG) and New Zealand White (NZW) rabbits. The population consisted of 363 F2 animals derived from 9 F1 bucks and 33 F1 does. 186 microsatellite and three SNP markers were informative for mapping. RESULTS: Out of 189 markers, which could be assigned to linkage groups, 110 markers were genetically mapped for the first time. The average marker distance was 7.8 cM. The map across all autosomes reached a total length of 1419 cM. The maternal linkage map was 1.4 times longer than the paternal. All linkage groups could be anchored to chromosomes. On the basis of the generated genetic map, we identified a highly significant QTL (genome-wide significance p < 0.01) for different carcass weights on chromosome 7 with a peak position at 91 cM (157 Mb), a significant QTL (p < 0.05) for bone mass on chromosome 9 at 61 cM (65 Mb), and another one for drip loss on chromosome 12 at 94 cM (128 Mb). Additional suggestive QTL were found on almost all chromosomes. Several genomic loci affecting the fore, intermediate and hind parts of the carcass were identified. The identified QTL explain between 2.5 to 14.6% of the phenotypic variance in the F2 population. CONCLUSIONS: The results present the most comprehensive genetic map and the first genome-wide QTL mapping study for carcass and meat quality traits in rabbits. The identified QTL, in particular the major QTL on chromosome 7, provide starting points for fine mapping and candidate gene search. The data contribute to linking physical and genetic information in the rabbit genome.

Convergent evolution of IL-6 in two leporids (Oryctolagus and Pentalagus) originated an extended protein.

Interleukin 6 (IL-6) is a class-I helical cytokine with a broad spectrum of biological activities and a gene structure conserved throughout vertebrates, with five coding exons. IL-6 from European rabbits belonging to the subspecies Oryctolagus cuniculus cuniculus was previously shown to differ from other mammals by extending an additional 27 amino acids. However, in other leporids (Sylvilagus spp and Lepus spp) that diverged from the European rabbit ~12 million years ago this mutation was not present. In this study, we extended the study of IL-6 for the Oryctolagus cuniculus algirus subspecies and five additional lagomorphs' genera (Brachylagus, Bunolagus, Pentalagus, Romerolagus, and Ochotona). We confirmed the presence of the mutated stop codon in both O. c. cuniculus and O. c. algirus. We found that the typical stop codon is present in Sylvilagus bachmani and Lepus europaeus, in agreement with previous reports, but also in Bunolagus, Brachylagus, and Ochotona. Remarkably, in Pentalagus we detected a deletion of the stop codon causing an extension of IL-6 for 17 extra residues. Our results indicate that the IL-6 extension in those species occurred by two independent events: one occurred between 2 and 8 million years ago in the ancestral of the Oryctolagus subspecies, and the other occurred in a Pentalagus ancestral at a maximum of 9 million years ago. The absence of this IL-6 extension in Bunolagus, sister genus of Oryctolagus, shows that this evolutionary event happened by convergence suggesting some functional relevance.

Spongy film of cellulosic polysaccharide as a dressing for aphthous stomatitis treatment in rabbits

To develop an experimental model of acute inflammation, like aphthous ulcers, in oral cavity of rabbits, and also, to evaluate the results of the application of a polysaccharide spongy film of molasses from sugar cane as assist treatment in the healing process. Twenty adult rabbits weighting between 2.5 kg and 3.9 kg were divided into two groups: experimental and control infected ulcers were induced on the jugal mucosa by surgical excision. They were treated at the experimental group by curettage and dressing with spongy film of cellulosic polysaccharide film, whereas saline solution was used in the control group. Temporal evolution of the healing area, histopathology and bacteriological analysis were used to evaluate the healing process on the 3rd (D3), 7th (D7) and 11th days (D11). The healing time and bacteriological study showed no statistical differences on the group means. Analyzing the histopathology of the experimental group we verified epithelial hyperplasia from D3 to D11, instead in the control group there was a greater clutter of the epithelial cells from the D3 to D11. The experimental model used caused aphthous ulcers and the polysaccharide sponge film can be used as an aid in the symptomatic treatment and healing of the ulcerative lesions of the oral mucosa.

Comparação entre dois protocolos de tratamento de ceratoconjuntivite seca experimentalmente induzida em coelhos / Comparison of two treatment protocols of experimentally induced keratoconjunctivitis sicca in rabbits

O objetivo deste estudo foi avaliar e comparar a eficácia de dois protocolos de tratamento de ceratoconjuntivite seca (CCS) experimentalmente induzida em coelhos: uma formulação oftálmica tópica composta por álcool polivinílico 1,4%, adicionado com acetilcisteína 10% e pilocarpina 1% (AAP), e outro protocolo com o uso do óleo de semente de linhaça (OL) tópico em forma de colírio, durante 12 semanas. Foram utilizados 15 coelhos machos, adultos, da raça Nova Zelândia, alocados aleatoriamente em três grupos: grupo C (controle), grupo AAP (formulação oftálmica) e grupo L (OL tópica). Os animais foram avaliados semanalmente pelo teste lacrimal de Schirmer, teste de fluoresceína e teste de Rosa Bengala; uma vez por mês, pelo exame de citologia esfoliativa ocular; ao final do experimento, pela análise histopatológica da córnea e conjuntiva. Os resultados demonstraram que houve um aumento maior na produção lacrimal quando utilizada a formulação oftálmica, e uma resolução mais rápida das úlceras de córnea, bem como diminuição no número de células desvitalizadas quando utilizado o óleo de semente de linhaça, além de aumento no número de células caliciformes em ambos os grupos de tratamento. A associação desses dois protocolos pode ser no futuro uma alternativa no tratamento da CCS.

The objective of this study was to evaluate and compare the effectiveness of two treatment protocol of experimentally induced keratoconjunctivitis sicca (KCS) in rabbits, a topical ophthalmic formulation composed by 1.4% povinilic alcohol added with 10% acetylcysteine and 1% pilocarpine (AAP) and another protocol with the topical use of the linseed seed oil (LO) in eye drop form f or 12 weeks. Fifteen male New Zealand white rabbits were aleatory allocated in 3 groups: Group C (Control), Group AAP (ophthalmic formulation) and Group L (LO topical). The animals were evaluated weekly using the Schirmer's tear test, fluorescein test and Rose Bengal test monthly for ocular cytology, and at the end of the experiment for histopathological analysis of cornea and conjunctive. The results demonstrated that there was a larger increase in the tear production when the ophthalmic formulation was us ed and a faster rapid resolution of corneal ulcers and decrease in the number of devitalized cells when linseed seed oil was used, besides an increase in the number of caliciform cells in both treatment groups. The association of those two protocols can be a future alternative in the treatment of KCS.

Avaliação macroscópica e microscópica de implante de Biomembrana em estômago de coelhos (Oryctolagus cuniculus) / Macroscopic and microscopic evaluation biomembrane implant in the stomach of rabbits (Oryctolagus cuniculus)

Neste estudo foi implantado um retalho de biomembrana de látex natural em substituição a um fragmento da parede de estômago de coelhos albinos, raça Nova Zelândia, adultos, machos não castrados (n=12), com o intuito de avaliar o processo de reparação tecidual no que se refere à biocompatibilidade, à capacidade de reparação tecidual e a possíveis complicações. Aos 15, 30 e 60 dias de pós-operatório, os animais foram sacrificados, mediante o emprego de tiopental sódico (200mg kg-1), para posterior avaliação macroscópica e análise histopatológica da interface do implante com o tecido nativo. Macroscopicamente, aos 15, 30 e 60 dias de pós-operatório observou-se presença de aderências na face serosa. Aos 60 dias de pós-operatório, em todos os animais avaliados, a biomembrana não foi encontrada. Sob microscopia de luz, aos 15 e 30 dias, verificaram-se descontinuidade das camadas muscular e mucosa, presença de infiltrado inflamatório polimorfonuclear. Foram visualizados vasos sanguíneos e fibras musculares. Aos 60 dias, as camadas mucosa, muscular estavam completamente reconstituídas. O implante foi biocompatível e forneceu arcabouço para orientação e desenvolvimento das camadas teciduais do estômago, mediante processos de reparação, restabelecendo a estrutura do órgão.

In this study, biomembrane of natural latex was utilized to replace a section of the stomach wall of New Zealand rabbits, adult and non-castrated males (n=12), in order to evaluate the tissue repair process in regards to its biocompatility, scar formation ability and possible complications. The animals were euthanized at fifteen, 30 and 60 days post operation, by use of sodium thiopental (200mg kg-1), followed by macroscopic and histopathological analysis of the implant interface with the native tissue. Macroscopically, at fifteen, 30, and 60 days post operation adherence was observed in the serosal wall. At 60 days post operation, the biomembrane is not in the stomach. Under light microscopy, at fifteen and 30 days, discontinuity of muscle layer and mucosa layer, and presence of polimorfonuclear population of inflammatory cells was observed. New vessels and muscle fibers were observed. At 60 days, the mucosa and muscle layers were complete reconstituted. The implants were biocompatible and had provided the mainframe for orientation and development of the tissue layers through repairing processes, thus reestablishing the organ structure.

The effect of simvastatin on the regeneration of surgical cavities in the femurs of rabbits

To evaluate the effect of a local application of simvastatin gel in repairing bone defects in the femurs of rabbits. METHODS: Two standard surgical cavities were created in the femoral epiphysis of 18 rabbits. In the simvastatin group (SG), the cavities were filled with a collagen sponge soaked in 0.5 ml of a simvastatin (1 mg) gel, and the cavities were covered with a biological membrane. The bone cavities in the second group (control group) were filled with a blood clot and covered with a biological membrane. On the 7th, 21st and 42nd days, six animals in each group were euthanized, and the femurs were subject to histological evaluation (vascularity, fibrosis, reactive bone formation, osteoblasts, and osteoclasts) and immunohistochemical (anti-VEGF and anti-osteocalcin) analysis. The results were analyzed using a Wilcoxon test (p<0.05). RESULTS: There were significant differences between the two groups: the SG had greater scores in comparison with the CG in terms of the degree of vascularity on the 7th and the 21st days, fibrosis on the 21st day, bone formation reaction on the 21st and the 42nd days and the number of osteoblasts and osteoclasts on the 42nd day. The immunohistochemical expression was also greater for osteocalcin and VEGF on the 7th, 21st and 42nd days. CONCLUSION: Surgical defects created in rabbit femurs were treated locally with simvastatin gel to stimulate bone repair, which promoted an ameliorative effect in the morphological and immunohistochemical markers of bone regeneration.

Evolution of viral sensing RIG-I-like receptor genes in Leporidae genera Oryctolagus, Sylvilagus, and Lepus.

One of the most severe European rabbit (Oryctolagus cuniculus) pathogens is myxoma virus (MYXV), a rabbit-specific leporipoxvirus that causes the highly lethal disease myxomatosis. Other leporid genera, Sylvilagus and Lepus, encompass species with variable susceptibilities to MYXV, but these do not develop the lethal form of the disease. The protective role of the retinoic acid-inducible gene-I (RIG-I/DDX58) in sensing MYXV in nonpermissive human myeloid cells prompted the study of the RIG-I-like receptor (RLR) family evolution in the three leporid genera. This viral-sensor family also includes the melanoma differentiation-associated factor 5 (MDA5/IFIH1), and the laboratory of genetics and physiology 2 (LGP2/DHX58). Considering specifically the MYXV susceptible host (European rabbit) and one of the virus natural long-term hosts (Sylvilagus bachmani, brush rabbit), the amino acid differences of positively selected sites in RIG-I between the two species were located in the protein region responsible for viral RNA recognition and binding, the repressor domain. Such differences might play a determinant role in how MYXV is sensed. When looking for episodic selection on MDA5 and LGP2 of the eastern cottontail (Sylvilagus floridanus), we also uncovered evidence of selective pressures that might be exerted by a species-specific leporipoxvirus, the Shope fibroma virus. Finally, a putative alternative splicing case was identified in Oryctolagus and Lepus MDA5 isoforms, corresponding to the deletion of one specific exon. This study provided the first insights into the evolution of the leporid RLR gene family that helps illuminate the origins of the species-specific innate responses to pathogens and more specifically to MYXV.

Clinical and histological effects of the temporary occlusion of the rabbit nasolacrimal duct and point using cyanoacrylate adhesives / Efeitos clínicos e histológicos da oclusão temporária do ponto e do duto nasolacrimais de coelhos com adesivos de cianoacrilato

The objective of this study was to evaluate the clinical and histological effects of occluding the nasolacrimal ducts and points of rabbits. For this study, 20 adult New Zealand rabbits, both males and females, weighing 3.2±0.4kg were allocated into two groups for n-butyl-cyanoacrylate occlusion (GB, n=10) or 2-octyl-cyanoacrylate occlusion (GO, n=10). The contralateral eyes served as the controls. The persistence of tears was evaluated daily using the Schirmer I test. Discomfort, eye discharge, epiphora, and conjunctival hyperemia were assessed prior to the procedure (T0) and during the 14 subsequent days (T1-T14). On days seven and 14, five animals from each group were euthanized, and their nasolacrimal ducts were collected, processed and analyzed by histology. In the GB group, the Schirmer test values differed from that at T0 at all of the subsequent time points, whereas there was no difference in the values observed from the GO group. Compared with the corresponding controls, the GO and GB groups differed significantly at almost all of the time points. When comparing the treatment groups, differences were found at T6, T7, T9, T10, T11, T12 and T14, with higher Schirmer values in the GB group. Epiphora was observed in the GB group from T1 to T8 and in the GO group from T1 to T6. Within seven days post-occlusion, histology revealed a moderate foreign body reaction, with marked necrosis and sloughing of the canalicular epithelium, in the GO group, which was absent at day 14. In the GB group, a marked inflammatory reaction and a mild foreign body reaction were found at day seven, and the foreign body reaction was prevalent at day 14. This study demonstrated that both adhesives were effective in obstructing the nasolacrimal ducts and points of rabbits and that their application and handling are easy and free of complications. However, both adhesives promoted inflammatory and foreign body reactions that evolved to repair and regeneration at day 14 of evaluation.

Avaliaram-se os efeitos clínicos e histológicos da oclusão do ponto e do duto nasolacrimais de coelhos. Para isso, utilizaram-se 20 coelhos adultos da raça Nova Zelândia, machos e fêmeas, com peso de 3,2±0,4kg, distribuídos em dois grupos: oclusão com n-butil cianoacrilato (GB, n=10) e oclusão com 2-octil cianoacrilato (GO, n=10). Os olhos contralaterais foram utilizados como controle. A permanência da lágrima foi avaliada diariamente pelo teste de Schirmer I. Foram avaliados o desconforto, secreção ocular, epífora e hiperemia conjuntival, previamente ao procedimento (T0) e durante 14 dias (T1-T14). Aos sete e aos 14 dias, cinco animais de cada grupo foram submetidos à eutanásia, e os dutos nasolacrimais, colhidos, processados e analisados à histologia. Em GB, os valores de Schirmer diferiram de T0 em todos os momentos; em GO não houve diferença. Na comparação com o respectivo controle, GB e GO diferiram significativamente em quase todos os momentos. Ao compararem-se os tratamentos, houve diferença em T6, T7, T9, T10, T11, T12 e T14, sendo os valores de Schirmer superiores em GB. Epífora esteve presente em GB de T1 a T8 e em GO de T1 a T6. À histologia, em GO, aos sete dias, notou-se moderada reação de corpo estranho com marcante necrose e descamação do epitélio canalicular; tais alterações estiveram ausentes aos 14 dias. Em GB, verificou-se, aos sete dias, acentuada reação inflamatória e discreta reação de corpo estranho; aos 14 dias, houve predomínio da reação de corpo estranho. Concluiu-se que ambos os adesivos foram eficazes na obstrução do ponto e do duto nasolacrimais de coelhos, sendo sua aplicação e manuseio fáceis e livres de intercorrências e ambos promoveram reação inflamatória e de corpo estranho que evoluíram para reparação e regeneração aos 14 dias de avaliação.

Morfometria do aparelho genital e resposta superovulatória de coelhas suplementadas com geleia real / Morphometry of the genital tract and superovulatory response in rabbits supplemented with royal jelly

Avaliou-se o efeito da suplementação com geleia real sobre a morfometria do aparelho genital, resposta superovulatória e qualidade embrionária de coelhas. Trinta e seis fêmeas foram distribuídas em quatro grupos (G), sendo: G1 (n=9) formado por animais não suplementados com geleia real, e G2, G3 e G4 (n=9 em cada grupo) por animais suplementados com 10, 20 e 40mg/dia de geleia real. A superovulação consistiu na aplicação de 40UI de gonadotrofina coriônica equina, seguida por 40UI de gonadotrofina coriônica humana, via intramuscular, 48 horas após, e submetidas à cobrição natural. Os animais foram sacrificados, e os embriões coletados 72 horas após a cópula. Não houve diferença estatística entre tratamentos para as variáveis analisadas. O peso médio do aparelho genital foi de 10,88±0,38g; dos ovários - direito e esquerdo -, 0,28±0,02g; e o índice gonadossomático, 0,02±0,0g. O número médio de estruturas totais recuperadas foi de 9,2±1,4; de embriões viáveis, 8,7±1,4; e de degenerados, 0,5±0,2. Dos embriões viáveis, 5,6±0,8 foram classificados como grau I; 2,3±0,5, como grau II; e 0,8±0,2, como grau III. A suplementação com geleia real na dose de até 40mg/dia não apresentou efeito estimulador sobre o aparelho genital e a qualidade embrionária de coelhas.

The purpose of this paper was to assess the effect of supplementation of royal jelly on the morphometry of the genital tract, superovulatory response and embryonic quality in adult rabbits. 36 rabbits were distributed in four groups (G), as follows: G1 (n=9): no supplementation with royal jelly, G2, 3 and 4 (n=9): supplementation with 10, 20 and 40mg/day of royal jelly. The superovulatory protocol consisted in the application of a dose of 40UI of Equine Chorionic Gonadotrophin, followed by a dose of UI of Human Chorionic Gonadotrophin, intramuscularly, 48 hours later, and submitted to natural cover. The animals were euthanasiated and the embryos collected 72 hours after copulation. There was no statistical difference for the analyzed variables (P>0.05). The average weight for the female genital tract was 10.89±0.38g; the average weight of right and left ovaries was 0.28±0.02g and the average gonadosomatic index rate was 0.02±0.0g. The number of total recovered structures was 9.2±1.4. The average number of viable embryos was 8.7±1.4 and of degenerated embryos was 0.5±0.2. Viable embryos were morphologically classified as degree I: 5.6±0.8; degree II: 2.3±0.5; degree III: 0.8±0.2. Royal jelly did not present stimulatory effect on organs related to the reproduction of rabbits, nor was it effective to improve their embryonic quality at dosages of up to 40mg of in natura royal jelly.

A systematic study of the brain base arteries in the rabbit (Oryctolagus cuniculus) / Sistematização das artérias da base do encéfalo e suas fontes de suprimento sanguíneo em coelho da raça Nova Zelândia (Oryctolagus cuniculus)

The brains of 30 New Zealand rabbits (Oryctolagus cuniculus) were injected with red stained latex. The arteries of the ventral surface of the brain were systematized on the right (R) and on the left (L) side with the respective percentage of appearance: the aortic arch emitted the braquicephalic trunk and the left subclavian artery (83.3%); or the braquicephalic trunk, the left common carotid artery and the left subclavian artery (16.7%). The braquicephalic trunk emitted the right and the left common carotid arteries and the right subclavian artery (83.3%); or the right common carotid artery and the right subclavian artery (16.7%). The common carotid arteries were divided into external and internal carotid arteries (96.7% on the R, 100% on the L.). The internal carotid artery to the R was present (96.7%) and absent (3.3%), and to the L, was present (100%). The rostral choroidal artery to the R was collateral branch of the rostral branch of the internal carotid artery (83.3%), collateral branch of caudal branch of the internal carotid artery (16.7%), and to the L was collateral branch of the rostral branch of the internal carotid artery (93.3%), collateral branch of the caudal branch of the internal carotid artery (6.7%). The middle cerebral artery to the R and to the L was single (80%) and double (20%). The rostral cerebral artery to the R had middle caliber (90%), thin caliber (6.7%) and too thin caliber (3.3%), and to the L had middle caliber (76.7%), thin caliber (16.7%) and too thin caliber (6.7%). The internal ethmoidal artery was absent (73.3%), present and single (26.7%). The caudal cerebral artery to the R was single (66.7%), double (26.7%) and triple (6.7%), and to the L was single (63.3%) and double (36.7%). The terminal branches of the right and left vertebral arteries were present (100%, and formed the basilar artery (100%). The ventral spinal artery was present (100%). The caudal cerebellar artery, to the R was single (43.3%), single with labyrinthic artery isolated (26.7%) and double (30%), and to the L was single (50%), single with labyrinthic artery isolated (6.7%), double (40%) and triple (3.3%). The trigeminal artery to the R and to the L was present (100%). The rostral cerebellar artery to the R was single (53.3%) and double (46,7%), and to the L was single (63.3%) and double (36.7%). The rabbit's cerebral arterial circle was caudally closed (100%) and rostrally closed (93.3%) or opened (6.7%). The brain was supplied by the vertebral-basilar and carotid systems.

Foram utilizados 30 encéfalos de coelhos Nova Zelândia (Oryctolagus cuniculus), injetados com látex, corado em vermelho, com objetivo de sistematizar as artérias da base do encéfalo e suas fontes de suprimento sanguíneo. Sistematizou-se a origem das fontes de suprimento sanguíneo para o encéfalo e as artérias (Aa) da face ventral do mesmo, tanto à direita (D) como à esquerda (E), com suas respectivas percentagens de aparecimento: o arco aórtico emitiu tronco braquiocefálico e artéria (A.) subclávia E (83,3%), ou tronco braquiocefálico, A. carótida comum E e A. subclávia E (16,7%). O tronco braquiocefálico lançou A. carótida comum D e E e A. subclávia D (83,3%), ou A. carótida comum D e A. subclávia D (16,7%). A. carótida comum dividiu-se em Aa carótidas externa e interna (96,7% D, 100% E). A. carótida interna D presente (96,7%) e ausente (3,3%), à E presente (100%). A. corióidea rostral D ramo colateral do ramo rostral da A. carótida interna D (83,3%), ramo colateral do ramo caudal da A. carótida interna D (16,7%), à E, ramo colateral do ramo rostral da A. carótida interna E (93,3%), ramo colateral do ramo caudal da A. carótida interna E (6,7%). A. cerebral média D e E ímpar (80%) e dupla (20%). A. cerebral rostral D com calibre médio (90%), calibre fino (6,7%), calibre muito fino (3,3%), à E, com calibre médio (76,7%), calibre fino (16,7%), calibre muito fino (6,7%). A. etmoidal interna ausente (73,3%), presente e ímpar (26,7%). A. cerebral caudal D, ímpar (66,7%), dupla (26,7%) e tripla (6,7%), à E, ímpar (63,3%) e dupla (36,7%). Ramos terminais --das Aa. vertebrais D e E presentes (100%) formaram a A. basilar (100%). A. espinhal ventral ímpar presente (100%). A. cerebelar caudal D, ímpar (43,3%), ímpar com A. labiríntica isolada (26,7%) e dupla (30%), à E, ímpar (50%), ímpar com A. labiríntica isolada (6,7%), dupla (40%) e tripla (3,3%). A. trigeminal D e E presente (100%). A. cerebelar rostral D, ímpar (53,3%) e dupla (46,7%), à E, ímpar (63,3%) e dupla (36,7%). Observou-se que o círculo arterial Cerebral do coelho foi fechado caudalmente (100%), rostralmente fechado (93,3%) e aberto (6,7%). O encéfalo foi suprido pelos sistemas vértebro-basilar e carotídeo.

Avaliação do efeito do alho (Allium sativum L. ) sobre o colesterol plasmático em coelhos com hipercolesterolemia induzida / Evaluation of garlic (Allium sativum L. ) effects on the serum cholesterol of rabbits with induced hypercholesterolemia

A utilização de plantas no tratamento de doenças ou como meio curativo é uma tradição popular e altamente difundida, sendo que muitos trabalhos abordam as propriedades terapêuticas e farmacológicas do alho na redução das hiperlipidemias. O objetivo deste estudo foi investigar a influência do extrato aquoso (E.A.) do alho (Allium sativum L.) no tratamento do colesterol plasmático em coelhos com hipercolesterolemia experimental. Os animais foram divididos em G1 (grupo controle) e G2 (grupo tratado com alho). O experimento foi desenvolvido em três fases: na 1ª fase os animais receberam dieta comercial de coelhos para avaliar o nível basal de colesterol nos animais; na 2ªfase, todos os animais passaram a receber dieta suplementada com gema de ovo, até o final do experimento, para desenvolver hipercolesterolemia, e; na 3ª fase os animais do grupo G2 receberam o tratamento com E. A. de alho. O colesterol na 1ª fase foi de 39,94 ± 9,57 mg dL-1. Na 2ª fase houve elevação significativa (p<0,05) no nível de colesterol plasmático nos dois grupos -acima de 100 mg dL-1. Com relação ao tratamento, o alho não promoveu redução no colesterol plasmático dos coelhos, contrapondo os dados da literatura.

The use of plants to treat diseases or even to cure them is a high diffused popular tradition, and several studies discuss the therapeutic and pharmacological properties of garlic in the reduction of hyperlipidaemias. The objective of the present study was to investigate the influence of the aqueous extract of garlic (Allium sativum) to treat the serum cholesterol of rabbits with experimental hypercholesterolemia. The animals were divided into G1 (control group) and G2 (group treated with garlic). The experiment was developed according to 3 phases: during the 1st phase, all animals were provided with a regular diet to evaluate the basal cholesterol; during the 2nd phase, all animals received a supplemented diet until the end of the experiment, in order to develop hypercholesterolemia and, in the 3rd phase, the animals on G2 received the garlic treatment. The cholesterol registered on the 1st phase was 39.94 ± 9.57 mg dL-1. On the 2nd phase, there was an increase on the serum cholesterol level in both groups - higher than 100 mg dL-1. Concerning the treatment, the garlic did not reduce the serum cholesterol in rabbits.

Pseudogenization of the MCP-2/CCL8 chemokine gene in European rabbit (genus Oryctolagus), but not in species of Cottontail rabbit (Sylvilagus) and Hare (Lepus).

BACKGROUND: Recent studies in human have highlighted the importance of the monocyte chemotactic proteins (MCP) in leukocyte trafficking and their effects in inflammatory processes, tumor progression, and HIV-1 infection. In European rabbit (Oryctolagus cuniculus) one of the prime MCP targets, the chemokine receptor CCR5 underwent a unique structural alteration. Until now, no homologue of MCP-2/CCL8a, MCP-3/CCL7 or MCP-4/CCL13 genes have been reported for this species. This is interesting, because at least the first two genes are expressed in most, if not all, mammals studied, and appear to be implicated in a variety of important chemokine ligand-receptor interactions. By assessing the Rabbit Whole Genome Sequence (WGS) data we have searched for orthologs of the mammalian genes of the MCP-Eotaxin cluster. RESULTS: We have localized the orthologs of these chemokine genes in the genome of European rabbit and compared them to those of leporid genera which do (i.e. Oryctolagus and Bunolagus) or do not share the CCR5 alteration with European rabbit (i.e. Lepus and Sylvilagus). Of the Rabbit orthologs of the CCL8, CCL7, and CCL13 genes only the last two were potentially functional, although showing some structural anomalies at the protein level. The ortholog of MCP-2/CCL8 appeared to be pseudogenized by deleterious nucleotide substitutions affecting exon1 and exon2. By analyzing both genomic and cDNA products, these studies were extended to wild specimens of four genera of the Leporidae family: Oryctolagus, Bunolagus, Lepus, and Sylvilagus. It appeared that the anomalies of the MCP-3/CCL7 and MCP-4/CCL13 proteins are shared among the different species of leporids. In contrast, whereas MCP-2/CCL8 was pseudogenized in every studied specimen of the Oryctolagus - Bunolagus lineage, this gene was intact in species of the Lepus - Sylvilagus lineage, and was, at least in Lepus, correctly transcribed. CONCLUSION: The biological function of a gene was often revealed in situations of dysfunction or gene loss. Infections with Myxoma virus (MYXV) tend to be fatal in European rabbit (genus Oryctolagus), while being harmless in Hares (genus Lepus) and benign in Cottontail rabbit (genus Sylvilagus), the natural hosts of the virus. This communication should stimulate research on a possible role of MCP-2/CCL8 in poxvirus related pathogenicity.

Copy number polymorphism in the α-globin gene cluster of European rabbit (Oryctolagus cuniculus).

Comparative genomic studies have revealed that mammals typically possess two or more tandemly duplicated copies of the α-globin (HBA) gene. The domestic rabbit represents an exception to this general rule, as this species was found to possess a single HBA gene. Previous electrophoretic surveys of HBA polymorphism in natural populations of the European rabbit (Oryctolagus cuniculus) revealed extensive geographic variation in the frequencies of three main electromorphs. The variation in frequency of two electromorphs is mainly partitioned between two distinct subspecies of European rabbit, and a third is restricted to the hybrid zone between the two rabbit subspecies in Iberia. Here we report the results of a survey of nucleotide polymorphism, which revealed HBA copy number polymorphism in Iberian populations of the European rabbit. By characterizing patterns of HBA polymorphism in populations from the native range of the European rabbit, we were able to identify the specific amino-acid substitutions that distinguish the previously characterized electromorphs. Within the hybrid zone, we observed the existence of a second HBA gene duplicate, named HBA2, that mostly represents a novel sequence haplotype, which occurs in higher frequency within the hybrid zone, and thus appears to have arisen in hybrids of the two distinct subspecies. Although this novel gene is also present in other wild Iberian populations, it is almost absent from French populations, which suggest a recent ancestry, associated with the establishment of the post-Pleistocene contact zone between the two European rabbit subspecies.