Management practices associated with strongylid parasite prevalence on horse farms in rural counties of Kentucky.

Anthelmintic resistance among cyathostomin parasites is a wide-spread problem. The parasite control guidelines written by the American Association of Equine Practitioners (AAEP) encourages the preservation of anthelmintic efficacy by reducing treatment frequency, using targeted deworming, and implementing environmental management practices. While there is knowledge regarding parasite management practices of affluent horse farms in the United States, surveys rarely explore the rural and underserved regions. The purpose of this study was to observe the management practices of horse farms in rural regions Kentucky, including working Amish farms, and determine factors associated with strongyle prevalence. A total of 160 horses among 38 owners from 28 different farms were enrolled in this study. A questionnaire survey regarding equine information, farm management, and deworming history was performed with each owner. Fecal samples were collected to determine fecal egg counts, perform coprocultures for subsequent strongyle larvae identification, and Strongylus vulgaris specific PCR. Serum samples were collected for the S. vulgaris antibody specific ELISA. The mean number of deworming treatments given in the last year was 2.1 with a 95% confidence interval of 1.9-2.3 with ivermectin being the most common active used. Statistical analysis showed horses treated within the last three months with a macrocylic lactone (ML) drug had significantly lower egg counts than horses treated with a ML 7-9 months ago (p = .0005). Despite the AAEP recommendations to reduce the overall number of treatments by using a surveillance-based approach and to no longer rotate treatments, only 17 horses reportedly had a fecal sample submitted for a fecal egg count and 65 horses were dewormed in a rotational manner. Horses whose owners utilized an informative deworming source (i.e., veterinarian, internet, magazine, local feed store) also had significantly lower counts (p = .0026). All coprocultures were negative for S. vulgaris while five horses were PCR positive. Interestingly, 95 horses tested ELISA positive for S. vulgaris. The strongyle egg counts of the working Amish horses were not significantly different from the other horses in this study and deworming practices including the use of efficacious drugs and low treatment frequencies were in accordance with the AAEP guidelines. This study was the first to summarize deworming management practices of rural regions in Kentucky, including a working Amish community. Overall, horse owners employed deworming practices recommended by the AAEP, however rotational deworming is still commonly implemented and fecal egg counts are rarely used.

Detection of Strongylus vulgaris in equine faecal samples by real-time PCR and larval culture - method comparison and occurrence assessment.

BACKGROUND: Strongylus vulgaris has become a rare parasite in Germany during the past 50 years due to the practice of frequent prophylactic anthelmintic therapy. To date, the emerging development of resistance in Cyathostominae and Parascaris spp. to numerous equine anthelmintics has changed deworming management and the frequency of anthelmintic usage. In this regard, reliable detection of parasitic infections, especially of the highly pathogenic S. vulgaris is essential. In the current study, two diagnostic methods for the detection of infections with S. vulgaris were compared and information on the occurrence of this parasite in German horses was gained. For this purpose, faecal samples of 501 horses were screened for S. vulgaris with real-time PCR and an additional larval culture was performed in samples of 278 horses. A subset of 26 horses underwent multiple follow-up examinations with both methods in order to evaluate both the persistence of S. vulgaris infections and the reproducibility of each diagnostic method. RESULTS: The real-time PCR revealed S. vulgaris-DNA in ten of 501 investigated equine samples (1.9%). The larval culture demonstrated larvae of S. vulgaris in three of the 278 samples (1.1%). A direct comparison of the two methods was possible in 321 samples including 43 follow-up examinations with the result of 11 S. vulgaris-positive samples by real-time PCR and 4 S. vulgaris-positive samples by larval culture. The McNemar's test (p-value = 0.016) revealed a significant difference and the kappa values (0.525) showed a moderate agreement between real-time PCR and larval culture. CONCLUSIONS: The real-time PCR detected a significantly higher proportion of positives of S. vulgaris compared to larval culture and should thus be considered as a routine diagnostic method for the detection of S. vulgaris in equine samples.

Attitudes towards implementation of surveillance-based parasite control on Kentucky Thoroughbred farms - Current strategies, awareness and willingness-to-pay.

REASONS FOR PERFORMING STUDY: Traditionally, equine parasite control has relied heavily on frequent routine anthelmintic treatments applied at regular intervals all year round. However, current recommendations aim to employ a more surveillance-based approach and it remains unknown to what extent these recommendations are being implemented on US horse farms. OBJECTIVES: To describe equine parasite control on Kentucky Thoroughbred farms and evaluate respondents' willingness to pay for various attributes of surveillance-based parasite control strategies. STUDY DESIGN: Questionnaire survey performed among the membership of the Kentucky Thoroughbred Farm Managers' Club. METHODS: The survey collected demographic data and information about current parasite control strategies. Further, respondents were asked to choose between hypothetical parasite control strategies described with a combination of different attributes: costs, time and effort needed, hypothetical disease-risk levels and hypothetical risks of anthelmintic resistance. Data were analysed with multivariable logistic analysis. RESULTS: About 26% responded to the survey (n = 112). Most respondents were concerned about anthelmintic resistance and incorporated veterinary advice in defining their deworming programme. However, almost 70% were following a traditional rotational deworming programme with little or no faecal surveillance. Respondents were willing to pay a premium for a product for which there is no known anthelmintic resistance and provided the highest possible decrease in health risks. The number of young horses on the farm, utilisation of veterinarian advice in developing a deworming programme, expressed concern about drug resistance in parasites and having documented drug resistance on the farm all associated significantly with the type of parasite control programme used. CONCLUSIONS: Traditional approaches for equine parasite control are still widely used in the Kentucky Thoroughbred industry. The data suggest that respondents were only willing to make these changes if they could be given assurance that the surveillance-based approach would prevent anthelmintic resistance and decrease health risks significantly for the horses.

Life cycle biological efficiency of mice divergently selected for heat loss.

Divergent selection in mice for heat loss was conducted in 3 independent replicates creating a high maintenance, high heat loss (MH) and low maintenance, low heat loss (ML) line and unselected control (MC). Improvement in feed efficiency was observed in ML mice due to a reduced maintenance energy requirement but there was also a slight decline in reproductive performance, survivability, and lean content, particularly when compared to MC animals. The objective of this study was to model a life cycle scenario similar to a livestock production system and calculate total inputs and outputs to estimate overall biological efficiency of these lines and determine if reduced feed intake resulted in improved life cycle efficiency. Feed intake, reproductive performance, growth, and body composition were recorded on 21 mating pairs from each line × replicate combination, cohabitated at 7 wk of age and maintained for up to 1 yr unless culled. Proportion of animals at each parity was calculated from survival rates estimated from previous research when enforcing a maximum of 4, 8, or 12 allowed parities. This parity distribution was then combined with values from previous studies to calculate inputs and outputs of mating pairs and offspring produced in a single cycle at equilibrium. Offspring output was defined as kilograms of lean output of offspring at 49 d. Offspring input was defined as megacalories of energy intake for growing offspring from 21 to 49 d. Parent output was defined as kilograms of lean output of culled parents. Parent input was defined as megacalories of energy intake for mating pairs from weaning of one parity to weaning of the next. Offspring output was greatest in MC mice due to superior BW and numbers weaned, while output was lowest in ML mice due to smaller litter sizes and lean content. Parent output did not differ substantially between lines but was greatest in MH mice due to poorer survival rates resulting in more culled animals. Input was greatest in MH and lowest for ML mice for both offspring and parent pairs, consistent with previous results in these lines. Life cycle efficiency was similar in MC and ML mice, while MH mice were least efficient. Ultimately, superior output in MC mice slightly outweighed the lower inputs in ML animals resulting from decreased maintenance energy requirements. Therefore, selection to reduce maintenance energy requirements may be more useful in terminal crosses or in a selection index to reduce possible negative effects on output, especially reproductive performance.

Body composition and feed intake of reproducing and growing mice divergently selected for heat loss.

Changes in maintenance energy requirements and in feed efficiency have been achieved by divergent selection for heat loss in mice in 3 replicates, creating high heat loss, high maintenance (MH) and low heat loss, low maintenance (ML) lines and an unselected control (MC). However, feed intake has mainly been measured in mature animals and not during growth or reproduction. Additionally, there is evidence that reducing maintenance energy will increase fat content, an undesirable result. To evaluate if selection has altered body composition and lifecycle feed intake, mating pairs were continuously mated and maintained for up to 1 yr unless culled. Offspring pairs were sampled from each line at each parity and maintained from 21 to 49 d of age. Feed intake was recorded for mating pairs throughout the year and on offspring pairs. Body weight was recorded on all animals at culling as well as percent fat, total fat, and total lean, measured by dual X-ray densitometry. Average daily gain was also recorded for offspring. Energy partitioning was achieved using 2 approaches: Approach I regressed energy intake of the pair on sum of daily metabolic weight and total gain to obtain maintenance (bm) and growth (bg) coefficients for each line, replicate, feeding period, and sex (offspring pairs only); Approach II calculated bm for each pair assuming constant energy values for lean and fat gain. Energy coefficients and body composition traits were evaluated for effect of selection (MH vs. ML) and asymmetry of selection ([MH + ML]/2 vs. MC). Both MC mating and offspring pairs tended to have greater BW than the average of the selection lines (P < 0.08). Males of offspring pairs weighed more than females (P < 0.01), while females of mating pairs weighed more than males (P < 0.01). Line was insignificant (P > 0.15) for body composition traits. Using Approach I, MH mice had a greater bm than ML mice for mating pairs (P = 0.03) but not offspring pairs (P = 0.50). For Approach II, MH had a greater bm than ML mice for both mating (P = 0.01) and offspring pairs (P = 0.01). The effect of selection for heat loss on body composition was smaller than previously reported and unlikely to outweigh the benefit of reduced feed intake, which was shown to be maintained throughout an entire lifecycle that included reproducing animals. Additionally, the reduction in energy intake seems primarily due to reduced maintenance energy costs, validating the success of the selection procedure.

Lifetime reproductive performance and survival analysis of mice divergently selected for heat loss.

Divergent selection for heat loss was implemented in mice creating maintenance high (MH) and low maintenance (ML) lines and an unselected control (MC) in 3 independent replicates. Mice from the ML line have improved feed efficiency, due to decreased maintenance energy requirement, but there is potential for a correlated decline in reproductive performance and survivability. Number fully formed (NFF), number born alive (NBA), number weaned (NW), litter weaning weight (LWW), pup weaning weight (PWW), fraction alive at birth (FAB), fraction alive at weaning, and birth interval were recorded at every parity on 21 mating pairs from each line × replicate combination cohabitated at 7 wk of age and maintained for up to 1 yr. Traits were summed over parities to evaluate lifetime production. Pairs were culled due to death or illness, no first parity by 42 d cohabitation, 2 consecutive litters with none born alive, 3 consecutive litters with none weaned, 42 d between parities, or average size of most recent 2 litters less than half the average of first 3 litters. Survival probabilities were produced and evaluated for each line and used to calculate mean number of parities using a Markov-chain algorithm assuming a maximum of 4, 6, 8, 10, or 12 parities or 1 yr. Line was insignificant for all litter traits while NFF, NW, and FAB decreased with parity (P < 0.05) and PWW tended to increase (P < 0.07). The MC mice had higher lifetime NW, LWW, and PWW (P < 0.04). Birth interval showed that MH mice had increasingly larger intervals while remaining the same in ML mice (P < 0.01). In the survival analysis, MC mice had the greatest survival rates overall, but ML mice had the greatest rates in the period up to 5 parities while MH mice had the greatest rates in later parities. This resulted in greater mean number of parities for ML mice up to maximum of 8 parities and higher means for MH mice when the maximum number of allowed parities was 10 or higher. Reproductive performance was not substantially affected by changing maintenance energy requirements. The ML animals appear to survive well in early parities and produce more parities when a low number of maximum parities is enforced, but this benefit declines in later parities and MH animals survive better and increase mean number of parities when turnover rates are low. Therefore, selection for low maintenance animals may be beneficial for systems desiring a short generation interval but less so for systems desiring longevity.

Equine parasite control under prescription-only conditions in Denmark--awareness, knowledge, perception, and strategies applied.

Due to widespread development of anthelmintic resistance in equine parasites, recommendations for their control are currently undergoing marked changes with a shift of emphasis toward more coprological surveillance and reduced treatment intensity. Denmark was the first nation to introduce prescription-only restrictions of anthelmintic drugs in 1999, but other European countries have implemented similar legislations over recent years. A questionnaire survey was performed in 2008 among Danish horse owners to provide a current status of practices and perceptions with relation to parasite control. Questions aimed at describing the current use of coprological surveillance and resulting anthelmintic treatment intensities, evaluating knowledge and perceptions about the importance of various attributes of parasite control, and assessing respondents' willingness to pay for advice and parasite surveillance services from their veterinarians. A total of 1060 respondents completed the questionnaire. A large majority of respondents (71.9%) were familiar with the concept of selective therapy. Results illustrated that the respondents' self-evaluation of their knowledge about parasites and their control associated significantly with their level of interest in the topic and their type of education (P<0.0001). The large majority of respondents either dewormed their horses twice a year and/or performed two fecal egg counts per horse per year. This approach was almost equally pronounced in foals, horses aged 1-3 years old, and adult horses. The respondents rated prevention of parasitic disease and prevention of drug resistance as the most important attributes, while cost and frequent fecal testing were rated least important. Respondents' actual spending on parasite control per horse in the previous year correlated significantly with the amount they declared themselves willing to spend (P<0.0001). However, 44.4% declared themselves willing to pay more than what they were spending. Altogether, results indicate that respondents were generally familiar with equine parasites and the concept of selective therapy, although there was some confusion over the terms small and large strongyles. They used a large degree of fecal surveillance in all age groups, with a majority of respondents sampling and/or treating around twice a year. Finally, respondents appeared willing to spend money on parasite control for their horses. It is of concern that the survey suggested that foals and young horses are treated in a manner very similar to adult horses, which is against current recommendations. Thus, the survey illustrates the importance of clear communication of guidelines for equine parasite control.

Hepatic mitochondrial efficiency in lines of mice differing in feed intake.

Selection was practiced for high (MH) and low (ML) heat loss using direct calorimetry to create lines of mice differing in feed intake per unit average BW (FI/BW). A total of 25 generations of selection was practiced during maintenance of an unselected control (MC). As a percentage of MC, mice of MH and ML lines differed in heat loss and FI/BW by 56 and 34%, respectively, when selection ceased. The purpose of this study was to determine hepatic mitochondrial efficiency in mice at maintenance across the genetic lines. After measurement of feed intakeand BW over a 2-wk period, mice were euthanized, and their livers were extracted. Livers were homogenized and mitochondria were isolated in buffer for measurement of oxygen consumption and, hence, mitochondrial activity using a Clark-type oxygen electrode. Mitochondria of ML mice expressed greater (P < 0.03) respiratory control ratio by 22% than that for MH mice, revealing greater degree of uncoupling during electron transport in MH mice. Difference in FI/BW (P < 0.001) was large with MH mice consuming 34% more than ML mice. The respiratory control ratio differed between mice of MH and ML lines as expected; however, regression analysis revealed that it explained essentially no variation in feed intake per BW.

Sustainable equine parasite control: perspectives and research needs.

Clinically important equine parasites are ubiquitous in managed horse populations. The traditional approach to parasite control is frequent administration of anthelmintics to all horses on a farm. However, increasing levels of anthelmintic resistance is forcing horse owners and veterinarians to shift this control paradigm. Treatment regimens involving routine deworming of all horses throughout the year are now being replaced by more sustainable approaches, which take in to account the importance of maintaining adequate parasite refugia. The selective therapy principle has been recommended for more than 15 years, but there is limited experience with this approach. The relative magnitude of the faecal egg count for an individual horse is a consistent trait, and this provides a reliable basis for selective therapy. But no studies have evaluated the consequences of selective therapy in the long-term, and such studies are strongly needed to validate this approach. Importantly, it remains unclear how selective therapy may affect the prevalence and intensity of other parasites of significant pathogenic potential (e.g. Strongylus vulgaris), which have become uncommon due to years of intensive chemotherapy. Consequently, a selective approach requires vigilant surveillance of the parasite fauna and intensity. This places a demands for reliable diagnostic tools. Also noteworthy is the fact that the majority of equine nematode parasites are more pathogenic during their larval stages, when they cannot be detected by traditional egg counting techniques. Consequently, parasite-specific diagnostic tools capable of assessing prepatent parasite burdens, and able to differentiate between strongyle species of different pathogenic potentials, would be of great value to the equine clinician. Tools for detecting infections with the tapeworm Anoplocephala perfoliata are laborious, difficult to interpret, and at present there is no established method to evaluate treatment efficacy. Thus, better diagnostic tools are needed for tapeworms as well. Biological control, especially the predacious fungi have demonstrated good potential as an adjunct for strongyle control and such a product could easily have a market in equine establishments. In summary, there is general agreement that the traditional treat-all at frequent interval approach should be abandoned, and that optimal parasite control can be maintained with far fewer anthelmintic treatments. But better diagnostic techniques and more evidence documenting the long-term consequences of selective therapy programs are needed to develop and validate systems for sustainable equine parasite control.

Differences in mitochondrial efficiency between lines of mice divergently selected for heat loss.

Divergent selection for heat loss was applied to lines of mice for 15 generations (G) in 3 replicates. Selection resumed at G42 and continued through G51 across all replicates. At the end of G51, differences in heat loss and feed intake per unit of BW were approximately 56 and 34%, respectively, between high heat loss (MH) and low heat loss (ML) lines, as a percentage of the control line (MC) mean. Rates of liver mitochondrial respiration states, degree of coupling, and mitochondrial efficiency were measured in G58 using a Clark-type oxygen electrode to investigate possible causes of underlying variation in maintenance requirements. Body composition, BW, liver weight, feed intake, and residual feed intake (RFI) were also measured or calculated. Results reported here represent data from 197 mature male mice from all replicates. There were no differences in BW (P = 0.91) between the selection lines. Selection had an effect on lean percentage (P = 0.02), with MH mice being leaner. Fat percentage differences between the selection lines tended toward significance (P = 0.13). Livers of MH mice were approximately 13% larger than livers of ML mice (P = 0.01). An effect of selection was observed (P < 0.01) in feed intake per unit BW, with MH mice consuming 29% more feed than ML mice in G58. Differences in state 2 and state 4 respiration rates were significant (P = 0.01), whereas state 3 rates approached significance (P = 0.06). Mitochondria of MH mice respired at a greater rate than mitochondria of ML mice in all states of respiration; ML mice had respiratory control ratios that were, on average, 8% greater than MH mice (P = 0.14). Although this difference only tended toward significance, we suspect a greater degree of coupling of mitochondrial processes exists in ML animals. Mice selected for reduced heat loss had ADP:oxygen ratios that were approximately 20% greater than MH mice (P = 0.03). Therefore, greater mitochondrial efficiency was expressed in the ML animals. Within a line-replicate, there was no correlation between ADP:O and feed intake per unit BW (P = 0.71). In addition, no correlation of ADP:O and RFI existed (P = 0.92). Although the selection lines differed in mitochondrial traits, including overall mitochondrial efficiency (ADP:oxygen), these differences were not a significant underlying cause of variation in feed intake per unit BW or in RFI estimates.

Renewed selection for heat loss in mice: direct responses and correlated responses in feed intake, body weight, litter size, and conception rate.

Divergent selection in mice was renewed in 3 independent replicates for high (MH) and low (ML) heat loss. An unselected control (MC) was maintained in all replicates. Heat loss was measured for individual male mice for 15 h, overnight in direct calorimeters. After 16 initial generations of selection followed by 26 generations of relaxed selection, divergent selection resumed for 9 generations. The realized selection applied was very close to the maximum possible selection according to the criteria and protocol. Selection differentials were greater for high than for low selection due to greater variation in the MH line. When corrected for SD, standardized selection differentials were similar for MH and ML selection. Unintended selection in MC was negligible. Realized heritability for divergence was 0.14 +/- 0.01, which was considerably less than that realized during the initial generations of selection (0.28 +/- 0.03). Realized heritabilities for MH selection (0.16 +/- 0.05) and for ML selection (0.07 +/- 0.06) were less, especially for ML selection, than were observed in the earlier generations. The difference in heat loss between MH and ML males was 55.7% of the MC mean at generation 51, compared with a difference of 53.6% in generation 15; this difference had decreased to 34.4% at the end of the relaxed selection (generation 42). For feed intake between 8 and 11 wk, MH and ML males differed by 34.0% of the MC mean by the end of the selection process. Body weight at 12 wk for MH and ML males was less than for MC males. Litter size response was positively related to the heat loss response. Conception rate was poorer in MH matings than in MC and ML matings.

Introducing biological phosphorus removal in an alternating plant by means of control: a full scale study.

In this paper, a control strategy for introducing enhanced biological phosphorus removal (EBPR) in an alternating plant designed for enhanced biological nitrogen removal (EBNR) is presented. Alternating aerobic and anaerobic conditions to promote EBPR are provided by controlling the phases of the operational cycle, instead of a separate anaerobic volume. By utilising the control schemes already built in the STAR control system for nitrogen removal, the control strategy is fully integrated in the system. The control system relies on on-line measurements of nitrogen (ammonia and/or nitrate) and orthophosphate. The control strategy has been implemented in full-scale operation at the Avedøre wastewater treatment plant in Denmark and the results show clear indications of success. The control strategy has operated robustly for several months with a 60% decrease in use of precipitation chemicals.

Comparison of feed energy costs of maintenance, lean deposition, and fat deposition in three lines of mice selected for heat loss.

Three replications of mouse selection populations for high heat loss (MH), low heat loss (ML), and a nonselected control (MC) were used to estimate the feed energy costs of maintenance and gain and to test whether selection had changed these costs. At 21 and 49 d of age, mice were weighed and subjected to dual x-ray densitometry measurement for prediction of body composition. At 21 d, mice were randomly assigned to an ad libitum, an 80% of ad libitum, or a 60% of ad libitum feeding group for 28-d collection of individual feed intake. Data were analyzed using 3 approaches. The first approach was an attempt to partition energy intake between costs for maintenance, fat deposition, and lean deposition for each replicate, sex, and line by multiple regression of feed intake on the sum of daily metabolic weight (kg(0.75)), fat gain, and lean gain. Approach II was a less restrictive attempt to partition energy intake between costs for maintenance and total gain for each replicate, sex, and line by multiple regression of feed intake on the sum of daily metabolic weight and total gain. Approach III used multiple regression on the entire data set with pooled regressions on fat and lean gains, and subclass regressions for maintenance. Contrasts were conducted to test the effect of selection (MH - ML) and asymmetry of selection [(MH + ML)/2 - MC] for the various energy costs. In approach I, there were no differences between lines for costs of maintenance, fat deposition, or protein deposition, but we question our ability to estimate these accurately. In approach II, selection changed both cost of maintenance (P = 0.03) and gain (P = 0.05); MH mice had greater per unit costs than ML mice for both. Asymmetry of the selection response was found in approach II for the cost of maintenance (P = 0.06). In approach III, the effect of selection (P < 0.01) contributed to differences in the maintenance cost, but asymmetry of selection (P > 0.17) was not evident. Sex effects were found for the cost of fat deposition (P = 0.02) in approach I and the cost of gain (P = 0.001) in approach II; females had a greater cost per unit than males. When costs per unit of fat and per unit of lean gain were assumed to be the same for both sexes (approach III), females had a somewhat greater estimate for maintenance cost (P = 0.10). We conclude that selection for heat loss has changed the costs for maintenance per unit size but probably not the costs for gain.

Correlated responses in maternal performance following divergent selection for heat loss in mice.

Divergent selection in mice was applied in 3 independent replicates for high (maintenance high; MH) and low (maintenance low; ML) heat loss for 16 generations. An unselected control (maintenance control; MC) was also maintained in all replicates. Selection ceased for 26 generations; heat-loss measurement and selection resumed at generation 42. Lactation performance, dam weight, dam feed intake, and efficiency of production of pup weight were recorded or calculated for MH and ML dams in all 3 replicates at generation 46 or 47 with the objective of determining whether selection for heat loss has created correlated responses in maternal performance. One-half of the dams reared their own litters, and one-half reared cross-fostered (across lines) litters. Between 10 and 12 litters were used from each replicate-line-rearing class. Litter size was recorded, and litters were standardized to 8 pups within 24 h of birth. For cross fostering, MH litters were matched to ML litters born within 24 h of each other, and MH-ML litter pairs were cross-fostered at 3 d of age. A weigh-suckle-weigh protocol was used to obtain milk production estimates over a 2-h suckling period at 6, 9, 12, and 15 d. Dam (plus litter) feed intake was also recorded at these times and was calculated as the disappearance of feed over 3-d intervals. Dams of the MH selection tended (P < 0.11) to have greater litter size than those of the ML selection; litter size of MC dams was intermediate. Line of dam affected milk production (P = 0.04) and dam feed intake (P < 0.03) as MH dams produced more milk and consumed more feed than ML dams. Average milk production for the 2-h measurement period was 1.70 +/- 0.07 and 1.41 +/- 0.07 g, and average 3-d feed consumption was 50.8 +/- 1.2 and 45.2 +/- 1.2 g for MH and ML dams, respectively. Cross-fostering had no effect (P > 0.86) on milk production. Line of dam tended to affect 21-d litter weight (P = 0.15) with litters reared by MH dams weighing more than those reared by ML dams, but there was no difference (P > 0.86) in 21-d dam weights. Efficiency of producing litter weight (litter 15-d weight: dam plus litter feed intake from d 6 to 15) was greater (0.49 vs. 0.46, SE = 0.009; P = 0.03) for ML than for MH dams. Selection for reduced heat loss (lower maintenance feed intake in the ML line) resulted in reduced milk production and feed intake in dams and greater efficiency of litter weight production.

Prescription-only anthelmintics--a questionnaire survey of strategies for surveillance and control of equine strongyles in Denmark.

In 1999, legislation in Denmark made anthelmintic drugs available only by prescription, and prohibited their use for routine, prophylactic treatment. A questionnaire survey was conducted in 2004 to determine current strategies for surveillance and control of equine strongyles used in Danish equine veterinary practices. Eighty-seven of 170 (51.2%) registered equine veterinary practices responded. Ninety seven percent of the respondents used faecal egg counts for diagnosis and surveillance, and 41% used larval cultures. Logistic regression revealed that the use of larval cultures was positively correlated with numbers of employees (P = 0.013) and the proportion of equine caseload in the practice (P < 0.000). Performing faecal egg counts and treating horses was most frequent during spring, summer and early autumn. Veterinary practices made treatment decisions based on cut-off values ranging from 20 to 500 eggs per gram (EPG) faeces. Ages of horses, clinical suspicions of parasitic disease, or requests by the owners were the most important factors in the strategies for faecal sampling. Less commonly, sampling strategies included all horses on the premises and random sampling. Ninety five percent of the respondents reported treating certain groups of horses without prior faecal analysis, including horses with clinical signs of parasitic disease (77%), foals (84%), horses less than 3 years of age (52%), and pregnant mares (51%). The respondents regarded the cyathostomin group as the most prevalent cause of parasitic disease and ill-thrift, followed by large strongyles and Parascaris equorum. Sixty seven percent rotated regularly between drugs, while 11% performed routine screening for anthelmintic resistance. Results of this survey suggest that limiting equine anthelmintics to prescription-only availability has increased the level of strongyle surveillance. Veterinary practitioners play a central role in equine parasite management with indications of a lowered intensity of treatment. However, screening for anthelmintic resistance remains uncommon.

Performance of mouse lines divergently selected for heat loss when exposed to different environmental temperatures. I. Reproductive performance, pup survival, and metabolic hormones.

Mouse populations differing in metabolic rate have been developed through selection for high (MH) and low (ML) heat loss, along with the unselected controls (MC). Objectives of the study were to compare the MH, ML, and MC lines for reproductive performance, pup survival, and metabolic hormones when reared at 12, 22, and 31 degrees C, and to search for line x environment interactions. Conception and litter size were recorded on the parent generation mice introduced to the environments at 11 wk of age and bred after a 3-wk acclimatization period. Survival of pups (preweaning to 3 wk; postweaning from 3 to 9 wk of age) was measured with continuous exposure in the designated environment from birth to the time of measurement. Corticosterone, triiodothyronine (T3), and thyroxine (T4) serum concentrations were measured on the parent generation after producing litters and on the pup generation at 9 wk. No line x environment interaction was detected for conception rate, preweaning mortality, postweaning survival, pup weaning weight, or body temperature. There were no differences in conception rate among lines and environments. Environments affected survival of pups, but there were no line differences. Rectal body temperatures were greater for MH than ML mice, and MC mice were intermediate; body temperature of mice did not differ among the environments. Lines differed significantly in litter size only in the 22 degrees C environment. No significant line differences were found for serum corticosterone or serum T3 or T4. Line x environment interaction was detected only for litter size and for serum corticosterone concentration in dams. Contrary to the other two lines, ML dam performance relative to MH and MC was not affected negatively by either of the thermal environments. Results from this study do not raise concern that selection to decrease maintenance requirements will produce livestock with any greater liability to cope and perform under an array of environmental temperatures.

Performance of mouse lines divergently selected for heat loss when exposed to different environmental temperatures. II. Feed intake, growth, fatness, and body organs.

Mouse populations differing in metabolic rate have been developed through selection for high (MH) and low (ML) heat loss, along with the unselected controls (MC). Objectives of the study were to compare the MH, ML, and MC lines for feed intake, growth, body fatness, and organ weights when reared at 12, 22, and 31 degrees C, and investigate potential line x environment interactions. Feed intake was recorded weekly from 3 to 9 wk of age, and BW at 3, 6, and 9 wk of age. Body fat percent and organ weights were measured at 9 wk of age. No line x environment interactions were detected for any of the traits measured. The MH mice consumed more feed than ML mice from 5 to 9 wk of age. Between 8 and 9 wk of age, MH mice consumed 13% more feed than the ML mice, but they were relatively leaner (14.45 vs. 16.32% body fat); MC mice were intermediate for both traits. Mice in the cold environment consumed the greatest amount of feed, and those in the hot environment consumed the least. Males consumed more feed than females, and the difference was greater in the cold than in the hot environment. No differences in BW were found between the lines. Mice in the 22 degrees C environment were heavier than their age-matched counterparts in the other two environments, and males were heavier than females at all ages. Relative to BW, the three lines had similar tail length, body length, and liver weight. Mice in the cold environment had heavier spleens and livers than those in the hot environment but relatively shorter bodies and tails; the normal environment was intermediate for these traits. Results from this study indicate that selection to decrease maintenance requirements did not produce mice with any less ability to grow and perform under an array of environmental temperatures.

Uncoupling proteins and energy expenditure in mice divergently selected for heat loss.

The objective of this study was to determine whether variation in energy expenditure created by selection on heat loss is mediated by uncoupling protein-1 (UCP1) in brown adipose tissue. Divergent selection for heat loss developed lines of mice with high (MH) and low (ML) maintenance energy expenditure. Concentration of UCP1 mRNA in brown adipose tissue (BAT) was 93% greater in ML than in MH mice (P < 0.02). Two new lines of mice, KH and KL, were bred by backcrossing a UCP1 knockout gene into the MH and ML lines, respectively; KH and KL with both knock-out (-/-) and wild type (+/+) UCP1 genotypes were generated. At 13 wk of age, KH mice exhibited greater heat loss (166 kcal x kg(0.75) x d(-1)) than KL mice (126.4 kcalkg(0.75) x d(-1)) regardless of the UCP1 knockout (P < 0.0001). Concentration of UCP2 mRNA in BAT was 74% greater in UCP1 knockout mice (-/-) than in wild type (+/+; P = 0.0001). We conclude that response to selection for increased energy expenditure was not mediated by increased expression or function of UCP1.

Locomotor activity, core body temperature, and circadian rhythms in mice selected for high or low heat loss.

Daily locomotor activity, core body temperature, and their circadian rhythms were measured in lines of mice selected for high (MH) or low (ML) heat loss and unselected controls (MC). Lines were created by selecting for 16 generations in each of three replicates. Collection of locomotor activity and core temperature data spanned Generations 20 and 21 for a total of 352 mice. Physical activity and core body temperature data were accumulated using implanted transmitters and continuous automated collection. Measurement for each animal was for 3 d. Activity was recorded for each half hour and then averaged for the day; temperature was averaged daily; circadian rhythm was expressed in 12-h (light vs dark) or 6-h periods as well as by fitting cyclic models. Activity means were transformed to log base 2 to lessen heterogeneity of variance within lines. Heat loss for a 15-h period beginning at 1630 and feed intake for 7 d were measured on 74 additional mice in order to estimate the relationship between locomotor activity and heat loss or feed intake. Selection lines were different (P < 0.01) for both locomotor activity and core body temperature. Differences were due to selection (MH-ML, P < 0.01), and there was no evidence of asymmetry of response (P > 0.38). Retransformed from log base 2 to the scale of measurement, mean activity counts were 308, 210, and 150 for MH, MC, and ML, respectively. Mean core temperatures were 37.2, 36.9, and 36.7 degrees C for MH, MC, and ML (P < 0.01), respectively. Females had greater physical activity (P < 0.01) and body temperature (P < 0.01) than males. There was no evidence of a sex x selection criterion interaction for either activity or temperature (P > 0.20). Overall phenotypic correlation between body temperature and log base 2 activity was 0.43 (P < 0.01). Periods during the day were different for both 12- and 6-h analyses (P < 0.01), but there were no period x selection criterion interactions (P > 0.1) for physical activity or body temperature. More sensitive cyclic models revealed significant (P < 0.01) 24-, 12-, 8-, and 6-h cycles that differed (P < 0.01) among lines. Estimated differences between MH and ML mice in feed intake and heat loss due to locomotor activity were 36 and 11.5%, respectively. Variation in activity thus contributed to variation in feed intake.

Gene expression in hypothalamus and brown adipose tissue of mice divergently selected for heat loss.

Gene expression was evaluated in mice divergently selected for 16 generations for heat loss, measured by direct calorimetry. The high (MH) heat loss line has approximately 50% greater heat loss, approximately 35% less body fat, approximately 20% greater feed intake, and twofold greater activity levels than the low (ML) heat loss line. At 11 wk, inbred males (developed from MH and ML) were euthanized 3 h after dark for dissection of tissues and extraction of RNA. Differential display PCR (DD-PCR) was used to evaluate transcriptional differences between lines in hypothalamus and brown adipose tissue (BAT). Evaluation was replicated within and across lines, using family pools of mRNA. Two genes were confirmed by competitive RT-PCR and/or Northern analysis to have greater levels of mRNA present in ML relative to MH mice. In both hypothalamus and BAT, the ribosomal protein L3 (RPL3) gene was expressed at higher levels in ML, whereas an unknown expressed sequence tag (EST) was also found at higher levels in the hypothalamus of ML mice. These results implicate RPL3 in regulation of energy balance and extend the genetic dissection of response to selection to the transcriptional level.