A simple geometric validation approach to assess the basic behaviour of space- and time- distributed models of epidemic spread - an example using the Ontario rabies model.

Dynamic mathematical modelling and stochastic simulation of disease-host systems for the purpose of epidemiological analysis offer great opportunities for testing hypotheses, especially when field experiments are impractical or when there is a need to evaluate multiple experimental scenarios. This, combined with the ever increasing computer power available to researchers, has contributed to the development of many mathematical models for epidemic simulations, such as the individual-based model (IBM). Nevertheless, few of these models undergo extensive validation and proper assessment of intrinsic variability. The Ontario rabies model (ORM) will be used here to exemplify some advantages of appropriate model behaviour validation and to illustrate the use of a simple geometric procedure for testing directional bias in distributed stochastic dynamic model of spread of diseases. Results were obtained through the comparison of 10 000 epizootics resulting from 100 epidemic simulations started using 100 distinct base populations. The analysis results demonstrated a significant directional bias in epidemic dispersion, which prompted further verification of the model code and the identification of a coding error, which was then corrected. Subsequent testing of the corrected code showed that the directional bias could no longer be detected. These results illustrate the importance of proper validation and the importance of sufficient knowledge of the model behaviour to ensure the results will not confound the objectives of the end-users.

Bioeconomic modelling of raccoon rabies spread management impacts in Quebec, Canada.

Beginning in 2006, point infection control operations and aerial distribution of oral rabies vaccines along the US border were performed in Quebec, Canada, to control the potential spread of raccoon rabies. A benefit-cost analysis assessed the economic efficiency of this rabies control programme into the future. In this study, a mathematical simulation model was used to determine the potential spread of raccoon rabies from the 2006 index case, and incidence rates of human post-exposure prophylaxis (PEP), animal testing and human exposure investigations were calculated. Benefits were calculated as the potential savings from reduced numbers of human PEP, animal testing and human exposure investigations owing to control, which ranged from $47 million to $53 million. Programme cost scenarios were based on projections of total expenditures, which ranged from $33 million to $49 million. Economic efficiency was indicated for approximately half of the modelled scenarios, with the greatest benefit-cost ratios resulting from reduced future programme costs.

Spatio-temporal variations and age effect on Toxoplasma gondii seroprevalence in seals from the Canadian Arctic.

Toxoplasmosis is a significant public health threat for Inuit in the Canadian Arctic. This study aimed to investigate arctic seals as a possible food-borne source of infection. Blood samples collected from 828 seals in 7 Canadian Arctic communities from 1999 to 2006 were tested for Toxoplasma gondii antibodies using a direct agglutination test. Polymerase chain reaction (PCR) was used to detect T. gondii DNA in tissues of a subsample of seals. Associations between seal age, sex, species, diet, community and year of capture, and serological test results were investigated by logistic regression. Overall seroprevalence was 10·4% (86/828). All tissues tested were negative by PCR. In ringed seals, seroprevalence was significantly higher in juveniles than in adults (odds ratio=2·44). Overall, seroprevalence varied amongst communities (P=0·0119) and by capture year (P=0·0001). Our study supports the hypothesis that consumption of raw seal meat is a significant source of infection for Inuit. This work raises many questions about the mechanism of transfer of this terrestrial parasite to the marine environment, the preponderance of infection in younger animals and the natural course of infection in seals. Further studies to address these questions are essential to fully understand the health risks for Inuit communities.

The analysis of crow population dynamics as a surveillance tool.

West Nile virus (WNV) infection, a zoonotic disease for which birds act as a reservoir, first appeared in North America in August 1999. It was first reported in Quebec in 2002. The Quebec surveillance system for WNV has several components, including the surveillance of mortality in corvid populations, which includes the American crow (Corvus brachyrhynchos). The main objectives of this study are to better understand the population dynamics of this species in Quebec and to evaluate the impact of WNV on these dynamics. We obtained observation data for living crows in this province for the period of 1990-2005 and then conducted a spectral analysis of these data. To study changes in crow population dynamics, the analysis was carried out before and after the appearance of WNV and space was divided in two different areas (urban and non-urban). Our results show the importance of cycles with periods of less than 1 year in non-urban areas and cycles with periods of greater than 1 year in urban areas in the normal population dynamics of the species. We obtained expected fluctuations in bird densities using an algorithm derived from spectral decomposition. When we compared these predictions with data observed after 2002, we found marked perturbations in population dynamics beginning in 2003 and lasting up to 2005. In the discussion, we present various hypotheses based on the behaviour of the American crow to explain the normal population dynamics observed in this species and the effect of type of area (urban versus non-urban). We also discuss how the predictive algorithm could be used as a disease surveillance tool and as a measure of the impact of a disease on wild fauna.

Projected effects of climate change on tick phenology and fitness of pathogens transmitted by the North American tick Ixodes scapularis.

Ixodes scapularis is the principal tick vector of the Lyme borreliosis agent Borrelia burgdorferi and other tick-borne zoonoses in northeastern North America. The degree of seasonal synchrony of nymphal and larval ticks may be important in influencing the basic reproductive number of the pathogens transmitted by I. scapularis. Because the seasonal phenology of tick vectors is partly controlled by ambient temperature, climate and climate change could shape the population biology of tick-borne pathogens. We used projected monthly normal temperatures, obtained from the second version of the Canadian Coupled Global Climate Model (CGCM2) under emissions scenario A2 of the Intergovernmental Panel on Climate Change for a site in southern Ontario, Canada, to simulate the phenology of I. scapularis in a mathematical model. The simulated seasonal abundance of ticks then determined transmission of three candidate pathogens amongst a population of white-footed mice (Peromyscus leucopus) using a susceptible-infected-recovered (SIR) model. Fitness of the different pathogens, in terms of resilience to changes in tick and rodent mortality, minima for infection duration, transmission efficiency and particularly any additional mortality of rodents specifically associated with infection, varied according to the seasonal pattern of immature tick activity, which was different under the temperature conditions projected for the 2020s, 2050s and 2080s. In each case, pathogens that were long-lived, highly transmissible and had little impact on rodent mortality rates were the fittest. However, under the seasonal tick activity patterns projected for the 2020s and 2050s, the fitness of pathogens that are shorter-lived, less efficiently transmitted, and more pathogenic to their natural hosts, increased. Therefore, climate change may affect the frequency and distribution of I. scapularis-borne pathogens and alter their evolutionary trajectories.

Role of migratory birds in introduction and range expansion of Ixodes scapularis ticks and of Borrelia burgdorferi and Anaplasma phagocytophilum in Canada.

During the spring in 2005 and 2006, 39,095 northward-migrating land birds were captured at 12 bird observatories in eastern Canada to investigate the role of migratory birds in northward range expansion of Lyme borreliosis, human granulocytic anaplasmosis, and their tick vector, Ixodes scapularis. The prevalence of birds carrying I. scapularis ticks (mostly nymphs) was 0.35% (95% confidence interval [CI] = 0.30 to 0.42), but a nested study by experienced observers suggested a more realistic infestation prevalence of 2.2% (95% CI = 1.18 to 3.73). The mean infestation intensity was 1.66 per bird. Overall, 15.4% of I. scapularis nymphs (95% CI = 10.7 to 20.9) were PCR positive for Borrelia burgdorferi, but only 8% (95% CI = 3.8 to 15.1) were positive when excluding nymphs collected at Long Point, Ontario, where B. burgdorferi is endemic. A wide range of ospC and rrs-rrl intergenic spacer alleles of B. burgdorferi were identified in infected ticks, including those associated with disseminated Lyme disease and alleles that are rare in the northeastern United States. Overall, 1.4[corrected]% (95% CI = 0.3 [corrected] to 0.41) of I. scapularis nymphs were PCR positive for Anaplasma phagocytophilum. We estimate that migratory birds disperse 50 million to 175 million I. scapularis ticks across Canada each spring, implicating migratory birds as possibly significant in I. scapularis range expansion in Canada. However, infrequent larvae and the low infection prevalence in ticks carried by the birds raise questions as to how B. burgdorferi and A. phagocytophilum become endemic in any tick populations established by bird-transported ticks.

Vector seasonality, host infection dynamics and fitness of pathogens transmitted by the tick Ixodes scapularis.

Fitness of tick-borne pathogens may be determined by the degree to which their infection dynamics in vertebrate hosts permits transmission cycles if infective and uninfected tick stages are active at different times of the year. To investigate this hypothesis we developed a simulation model that integrates the transmission pattern imposed by seasonally asynchronous nymphal and larval Ixodes scapularis ticks in northeastern North America, with a model of infection in white-footed mice (Peromyscus leucopus) reservoir hosts, using the bacteria Borrelia burgdorferi and Anaplasma phagocytophilum as examples. In simulations, survival of microparasites, their sensitivity to reduced rodent and tick abundance, and to 'dilution' by a reservoir-incompetent host depended on traits that allowed (i) highly efficient transmission from acutely-infected hosts, (ii) long-lived acute or 'carrier' host infections, and/or (iii) transmission amongst co-feeding ticks. Minimum values for transmission efficiency to ticks, and duration of host infectivity, necessary for microparasite persistence, were always higher when nymphal and larval ticks were seasonally asynchronous than when these instars were synchronous. Thus, traits influencing duration of host infectivity, transmission efficiency to ticks and co-feeding transmission are likely to be dominant determinants of fitness in I. scapularis-borne microparasites in northeastern North America due to abiotic forcings influencing I. scapularis seasonality.

Network analysis of Danish cattle industry trade patterns as an evaluation of risk potential for disease spread.

Trade patterns of animal movements in a specific industry are complex and difficult to study because there are many stakeholders, premises that are heterogeneously spread over the country, and a highly dynamic flow of animals exists among them. The Danish cattle industry was defined as a network of animal movements and graph theory was used to analyse the movements of cattle within this network. A premise was defined as a farm, an abattoir or a market. These premises constituted the network nodes in the graph and the animal movements between them were the links. In this framework, each premise had a sub-network of other premises to which it was linked by these animal movements. If no movement of animals were registered for a specific farm, then the sub-network for that premise consisted of only that premise. Otherwise, the sub-network linked the premise of interest to all premises from which and to which animals were moved, as long as there was a path linking animal movements to that specific premise. This approach allowed visualization and analyses of four levels of organization that existed in Denmark animal registers: (1) the animal that was moved, (2) the movements of all animals between two premises, (3) the specific premise network, and (4) the overall industry network. When contagious animals are moved from one premise to another, then to a third and so forth, these movements create a path for potential transfer of pathogens. The paths within which pathogens are present identify the transmission risks. A network of animal movements should provide information about pathogen transmission and disease spread. The network of the Danish cattle industry network was a directed scale-free graph (the direction of a movement was known), with an in-degree power of 2 an out-degree power of 1.46, consisted of 29,999 nodes, and 130,265 movements during a 6-month period. The in clustering coefficient was calculated to be 0.52 for the inward direction (movement to), while it was 0.02 for the outward direction (movement from). In Denmark, the cattle movements between premises demonstrated a large degree of heterogeneity. This heterogeneity in movements between farms should be used to evaluate the risk potential of disease transmission for each premise and must be considered when modelling disease spread between premises. The objective of this research was to describe the network of animal movements and not just the animal movements per se.

Climate change and the potential for range expansion of the Lyme disease vector Ixodes scapularis in Canada.

We used an Ixodes scapularis population model to investigate potential northward spread of the tick associated with climate change. Annual degree-days >0 degrees C limits for I. scapularis establishment, obtained from tick population model simulations, were mapped using temperatures projected for the 2020s, 2050s and 2080s by two Global Climate Models (the Canadian CGCM2 and the UK HadCM3) for two greenhouse gas emission scenario enforcings 'A2'and 'B2' of the Intergovernmental Panel on Climate Change. Under scenario 'A2' using either climate model, the theoretical range for I. scapularis establishment moved northwards by approximately 200 km by the 2020s and 1000 km by the 2080s. Reductions in emissions (scenario 'B2') had little effect on projected range expansion up to the 2050s, but the range expansion projected to occur between the 2050s and 2080s was less than that under scenario 'A2'. When the tick population model was driven by projected annual temperature cycles (obtained using CGCM2 under scenario 'A2'), tick abundance almost doubled by the 2020s at the current northern limit of I. scapularis, suggesting that the threshold numbers of immigrating ticks needed to establish new populations will fall during the coming decades. The projected degrees of theoretical range expansion and increased tick survival by the 2020s, suggest that actual range expansion of I. scapularis may be detectable within the next two decades. Seasonal tick activity under climate change scenarios was consistent with maintenance of endemic cycles of the Lyme disease agent in newly established tick populations. The geographic range of I. scapularis-borne zoonoses may, therefore, expand significantly northwards as a consequence of climate change this century.

A dynamic population model to investigate effects of climate on geographic range and seasonality of the tick Ixodes scapularis.

A dynamic population model of Ixodes scapularis, the vector of a number of tick-borne zoonoses in North America, was developed to simulate effects of temperature on tick survival and seasonality. Tick development rates were modelled as temperature-dependent time delays, calculated using mean monthly normal temperature data from specific meteorological stations. Temperature also influenced host-finding success in the model. Using data from stations near endemic populations of I. scapularis, the model reached repeatable, stable, cyclical equilibria with seasonal activity of different instars being very close to that observed in the field. In simulations run using data from meteorological stations in central and eastern Canada, the maximum equilibrium numbers of ticks declined the further north was the station location, and simulated populations died out at more northerly stations. Tick die-out at northern latitudes was due to a steady increase in mortality of all life stages with decreasing temperature rather than a specific threshold event in phenology of one life stage. By linear regression we investigated mean annual numbers of degree-days >0 degrees C (DD>0 degrees C) as a readily mapped index of the temperature conditions at the meteorological stations providing temperature data for the model. Maximum numbers of ticks at equilibrium were strongly associated with the mean DD>0 degrees C (r2>0.96, P<0.001), when the Province of origin of the meteorological station was accounted for (Quebec>Ontario, beta=103, P<0.001). The intercepts of the regression models provided theoretical limits for the establishment of I. scapularis in Canada. Maps of these limits suggested that the range of southeast Canada where temperature conditions are currently suitable for the tick, is much wider than the existing distribution of I. scapularis, implying that there is potential for spread. Future applications of the model in investigating climate change effects on I. scapularis are discussed.

Factors affecting milk urea nitrogen and protein concentrations in Quebec dairy cows.

Milk urea nitrogen (MUN), together with percentage milk protein (PROT), are increasingly used as indicators of the protein-energy balance and for monitoring nutrition and diagnosing feeding disorders. The goal of this study was to analyze the effects of parity, milk yield, days in lactation, somatic-cell count, and herd and feeding factors on MUN and PROT. In 10 dairy herds, one milk sample was taken from each of 418 cows, within +/- 2 days of the routine milk-test visit. We used a four-step multiple linear-regression model with backward elimination, including interactions between herd and the different factors. For both dependent variables, there were significant interactions with herd. Herd-specific models were markedly different; however, the daily amount of protein concentrates fed remained (and had a positive coefficient) in seven of 10 herd models for MUN. This factor is easy to record under field conditions and has to be considered in the evaluation of the ration by means of MUN and PROT. Overfeeding of rumen soluble protein can be easily diagnosed and corrected using MUN analyses. The relationships between MUN and PROT in respect of the factors parity, daily milk yield, and days postpartum also vary considerably among herds.

An epidemiological study of calcium metabolism in non-paretic postparturient Holstein cows.

Data from 1021 lactations of non-paretic Holstein cows followed in 14 Québec dairy herds were used to describe calcium 'metabolism' after calving in healthy cows. Serum total calcium, phosphorus, magnesium, potassium, albumin, and glucose were measured on the first and seventh days post-calving. The distributions were described and compared between the first and seventh day postpartum. The relationships between serum calcium on the one hand and the other serum metabolites and the cow's age on the other hand were assessed using a general linear model. Serum calcium and phosphorus values were lower on the first day postpartum than a week later (2.03 +/- 0.26 vs 2.26 +/- 0.18 mmol/l, 1.78 +/- 0.48 vs 1.93 +/- 0.39 mmol/l, respectively), whereas it was the opposite for glucose, magnesium, and potassium (3.98 +/- 0.95 vs 3.12 +/- 0.60 mmol/l, 1.01 +/- 0.35 vs 0.95 +/- 0.13 mmol/l, 4.84 +/- 0.40 vs 4.69 +/- 0.38 mmol/l, respectively). Albumin values were similar (25.7 +/- 3.3 vs 25.2 +/- 3.4 g/l). On the first day postpartum, serum calcium was associated in a curvilinear fashion with age, phosphorus and albumin. It also was associated, but to a lesser extent, with glucose and magnesium, whereas it was not associated with potassium. On the seventh day postpartum, calcium was associated with age, phosphorus and with an increased importance of albumin. The results are discussed with regard to postpartum hypocalcemia, the interpretation of serum metabolite values after calving, and the use of the physiological stress at calving. We concluded that (1) postpartum hypocalcemia was an event to be expected, especially for the older cow, (2) a multivariable approach should be used to interpret biochemical profiles after calving, and (3) such profiles could be used to better assess the cow's health.

Psychodemographic profile of stockpeople working on independent and integrated swine breeding farms in Quebec.

As a part of a larger study, the psychodemographic profile of Quebec pig stockpeople was described based on a sample of 48 persons working on independent farms randomly selected in the Richelieu-Saint-Hyacinthe region, and on a convenience sample of 38 others employed by 5 organizations of integrated swine production chosen among the largest ones in Québec. The 2 groups of stockpeople were described separately because of the differences between the 2 types of production. Demographic data (age, sex, level of education, training and experience in swine production, seniority on the farm) were obtained through a face-to-face interview. The personality profile was assessed using a standard personality test (French translation of the Sixteen Personality Factor Questionnaire). Both the stockpersons on independent farms and those employed by the organizations were more reserved, emotionally stable, serious, conscientious, unsentimental, controlled, introverted, and less anxious than the average person in the general population. However, multivariate analyses revealed some differences between these two groups of stock-persons (P = 0.021) as well as between the employees of the different organizations (P = 0.0038). These differences in personality profiles may reflect differences in working conditions, particularly regarding the human relationships, and in corporate cultures of the organizations.

Characterization of Serpulina hyodysenteriae isolates of serotypes 8 and 9 by random amplification of polymorphic DNA analysis.

A PCR-based DNA fingerprinting method termed RAPD (Random Amplification of Polymorphic DNA), or AP-PCR (for Arbitrary Primed PCR) was used to detect sequence diversity among reference strains and isolates of Serpulina hyodysenteriae. RAPD fingerprinting of 20 S. hyodysenteriae isolates of serotypes 8 or 9 from Quebec was generated with 2 different 10-base primers used independently. Reference strains and field isolates belonging to serotypes 8 or 9 revealed polymorphisms in RAPD fingerprints with both primers. Interspecies polymorphisms were observed by RAPD analysis of S. hyodysenteriae representing serotypes 1 to 9, S.innocens, and 5 other weakly beta-hemolytic intestinal spirochetes. A dendrogram based on the analysis of RAPD profiles of the strains tested with one of the primers (#17), permitted the clustering of these strains into 11 divisions. The predominance of particular RAPD profiles among S. hyodysenteriae isolates isolated from cases of swine dysentery in different herds suggested that certain S. hyodysenteriae types could be epidemiologically important. Our results indicate that RAPD could be used as a typing method for S. hyodysenteriae and as an epidemiological method for identifying spirochetes isolated from swine.

Survey of management and housing in farrowing quarters among independent and integrated swine farms in Québec.

Forty-eight randomly selected owner-operated swine breeding farms (independent farms) and 38 belonging to 5 integrated organizations specializing in swine production chosen from the largest in the province of Québec (integrated farms) were separately described regarding their general characteristics, sow feeding, management practises, and housing features in farrowing quarters. The parallel description of these 2 groups of farms aids in understanding what is done in the field. It also provides insight into potential differences between independent and integrated farms. Generally speaking, production tended to be more specialized and concentrated in integrated organizations. Specifically, more new practises seemed to have been adopted on the integrated farms, and their stockpersons seemed to have a more proactive style of management in farrowing quarters. Increased size of operations, proximity of information sources, profits yielded by new practises, and ease of implementation are discussed as explanations for this higher rate of adoption of new techniques among the organizations. These differences between the independent farms and the integrated organizations appeared to be all related to basic differences in their respective sizes. Although some differences were observed within, as well as between, each organization, many similarities were found across the majority of farms within each organization, thus supporting the existence of policies specific to each organization. Although these findings have to be confirmed before being generalized, they tend to suggest that independent swine farms and integrated organizations should be considered differently.

Detection of cervical bacterial contamination in swine by two methods of swabbing in relation to artificial insemination.

Factors associated with uterine contamination during artificial insemination (AI) are not well defined. A frequently imputed risk factor is vulvar hygiene, although its role has never been assessed objectively. The aim of this study was to identify an objective marker of hygiene during AI and to assess the impact of vulvar hygiene on cervical contamination. In a herd in a depopulation-repopulation process, 68 paired sows of each parity were divided into 2 treatment groups. Before 2 sham AIs, with a 24 hour interval, control sows (CTR) had their vulva cleaned and treatment sows (TRT) had theirs soiled with feces. After the 2nd sham AI, swabbings were taken from the spirette and from the cervix. Bacterial growth was assessed by a semiquantitative method, and aerobic bacterial species identified. The discordance between the paired data was assessed by a McNemar chi-square test. No difference in gram-positive bacterial counts between the 2 groups was found using the cervical swab (P > 0.05). The presence of trace colonies of Escherichia coli were, however, more frequent in TRT (P < 0.05). The spirette showed a greater contamination for mixed flora of bacterial species such as E. coli (P < 0.001) and non-S. suis streptococci (P < 0.05) in TRT sows. A light cervical E. coli growth was more frequent when the vulva had been soiled. Bacterial flora on the spirette following AI may be used as an objective and practical indicator of vulvar hygiene during AI in sows.

Importance of Escherichia coli in young beef calves from northwestern Quebec.

The objectives of this study were: (i) to investigate the prevalence of Escherichia coli producing F5 (K99), F41, or F165 fimbriae and STa enterotoxin; (ii) to determine serum antibody levels against these fimbriae; (iii) and to examine the association between bacteriological and serological results and the presence of diarrhea, in beef calves from northwestern Quebec. A total of 373 live three to four week old calves and 27 dead calves were sampled between January and March 1991. No isolates positive for F5 were detected in live calves, and only one E. coli producing STa and F41 was isolated. Escherichia coli producing F41-like surface antigens or F165 fimbriae were isolated from 17.43% and 5.63% of live calves, respectively. Antibodies against F5, F41 and F165 were low. Escherichia coli isolates positive for F41-like surface antigen were most often observed in calves born between January and March. No association was found between bacteriological and serological findings, nor between these findings and diarrhea. Calves born from dams vaccinated against E. coli had higher median antibody levels than those born from unvaccinated dams. No E. coli positive for F5 or F41 fimbriae were isolated from dead calves. Escherichia coli with F41-like surface antigen or F165 were found in 55.56% and 11.11% of ileal samples; 4% and 16% of cecal samples, and 0% and 7.4% of colon samples, respectively. Escherichia coli positive for F41-like surface antigen were detected significantly more frequently in the ileum (chi (2)2df = 31.01, p < 0.001).

A seroepidemiological study of the importance in cow-calf pairs of respiratory and enteric viruses in beef operations from northwestern Quebec.

Serum antibody analyses for bovine herpesvirus type 1 (BHV-1), bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine coronavirus (BCV), and bovine rotavirus (BRV) were performed on 527 randomly selected cows, before calving, and on 407 three-week-old calves. In cows and calves, BCV and BRV were the most seroprevalent viruses (80% to 100% according to virus and vaccination status). Bovine respiratory syncytial virus was the least seroprevalent in the cows, independent of the vaccination status. In nonvaccinated cows the seroprevalence to BRSV was 36.7%, and 53.5% in cows vaccinated less than two weeks prior to collecting blood, and 67.6% in cows vaccinated two weeks or more prior to blood collection. In their calves, BHV-1 was the least seroprevalent, independent of the vaccination status. The serological status and antibody titers in calves were generally associated with those of the dam. The occurrence of respiratory diseases in the calves was associated with cow and calf serological profiles (BHV-1, BRSV and BCV in the nonvaccinated group, BHV-1, BVDV and BCV in the vaccinated group). The occurrence of diarrhea was not associated with cow and calf serological profiles but was negatively associated with high level calf serum IgG in the nonvaccinated group (odds ratio = 0.73). Bovine coronavirus and BRV were shed by 1.4% and 4.9% of calves in the nonvaccinated group, and by 0% and 9.9% of calves in the vaccinated group, respectively. Bovine rotavirus shedding was associated with fecal diarrheic consistency at the moment of fecal sampling but not with previous occurrence of diarrhea.

In vitro viability of cryopreserved equine embryos following different freezing protocols.

The main objective of this study was to evaluate two freezing protocols and the effect of agar embedding on survival of day 6.5 equine embryos. A total of 133 embryos were used, in one group (n = 51), embryos were first embedded in agar before the freezing protocol was started. A freezing protocol to -30 degrees C or -33 degrees C was used before plunging embryos into liquid nitrogen (LN2). The embryos were thawed in water at 37 degrees C, evaluated and placed in culture. After 24 h culture, the embryos were evaluated for their morphology and development. No differences were observed between embryos plunged at -30 degrees or at -33 degrees C in LN2. The analysis of the morphology and development after thawing showed that the diameter and developmental stage at freezing correlated with embryo survival. Morula and early blastocyst stages of development were associated with better quality after freezing and thawing and had a better potential to survive after in vitro culture (p < 0.05) compared to more advanced stages. The agar failed to protect embryos from zona pellucida damage, but a tendency to prevent rupture was observed in larger embedded embryos.

Genomic relatedness among reference strains of different Streptococcus suis serotypes.

Hybridization studies using genomic DNA and a rDNA probe revealed genetic relatedness among reference strains of different Streptococcus suis serotypes. Although most serotype 22 isolates are biochemically atypical, the reference strain of capsular type 22 is genetically related to other S. suis serotypes, but not to Streptococcus pneumoniae. Using DNA digested with BamHI and BglII for ribotyping, some S. suis reference strains had common patterns, but this analysis mainly revealed variations in patterns of S. suis strains of different serotypes.