Effect of estrogen and progesterone on intracellular free zinc and zinc transporter expression in bovine oviduct epithelial cells.

Zinc (Zn) plays essential roles in numerous cellular processes. However, there is limited understanding of Zn homeostasis within the bovine reproductive system. This study investigated the influence of estradiol (E2) and progesterone (P4) on Zn transporter expression and intracellular free Zn levels in bovine oviduct epithelial cells (BOEC). For this purpose, cells were harvested from slaughtered cows and cultured in vitro. Intracellular Zn concentrations were measured using FluoZin-3AM staining, while real-time polymerase chain reaction assessed Zn transporter gene expression and quantification. Overall, our results confirmed the gene expression of all the evaluated Zn transporters (ZIP6, ZIP8, ZIP14, ZnT3, ZnT7 and ZnT9), denoted and the active role of E2 and P4 in intracellular Zn regulation. Our findings suggest an interaction between Zn, E2 and P4.

Effect of follicular wave synchronization using a progesterone-releasing intravaginal device-based protocol on in vitro embryo production in Bos taurus cows subjected to 14-day intervals ovum pick-up.

Follicular wave synchronization (FWS) before ovum pick-up (OPU) is one of the strategies used to improve the efficiency of in vitro embryo production (IVP). This study aimed to evaluate the effect of FWS on the total follicular number, cumulus-oocyte complex (COC) recovery, and in vitro embryo development in Angus cows (n = 33) subjected to OPU with 14-day intersession intervals. Additionally, it was also evaluated the presence of carryover effects given the short intersession interval used. The experiment was run as a 2-treatment (FWS vs. Control) x 2-period (1 vs. 2) crossover design. Animals in the FWS group received an intravaginal progesterone implant (1gr), estradiol benzoate (2 mg), and D-cloprostenol (150 µg) on day 0 and the OPU was performed on day 5. Control group animals did not receive any hormone treatment. The FWS increased the number of 6-10 mm follicles (P = 0.05), but it decreased the COC recovery rate (P < 0.01). The FWS did not affect the total or frozen embryo numbers (P = 0.49 and P = 0.17; respectively), but it increased the total blastocyst cell number (P < 0.01). A carryover effect was found on the total and < 6 mm follicles number (P = 0.10 and P < 0.01; respectively), and on the regular, atretic, viable, and total number of COC (P = 0.01, P = 0.08, P = 0.02 and P < 0.01; respectively). We concluded that the FWS increased the quality of embryos after OPU with 14-day intersession intervals in Angus cows and that this kind of OPU/IVP scheme enabled the existence of a carryover effect, especially on the follicle number and COC morphology.

Monitoring of Body Condition in Dairy Cows to Assess Disease Risk at the Individual and Herd Level.

A retrospective longitudinal study assessing the explanatory and predictive capacity of body condition score (BCS) in dairy cows on disease risk at the individual and herd level was carried out. Data from two commercial grazing herds from the Argentinean Pampa were gathered (Herd A = 2100 and herd B = 2600 milking cows per year) for 4 years. Logistic models were used to assess the association of BCS indicators with the odds for anestrus at the cow and herd level. Population attributable fraction (AFP) was estimated to assess the anestrus rate due to BCS indicators. We found that anestrus risk decreased in cows calving with BCS ≥ 3 and losing ≤ 0.5 (OR: 0.07-0.41), and that anestrus rate decreased in cohorts with a high frequency of cows with proper BCS (OR: 0.22-0.45). Despite aggregated data having a good explanatory power, their predictive capacity for anestrus rate at the herd level is poor (AUC: 0.574-0.679). The AFP varied along the study in both herds and tended to decrease every time the anestrous rate peaked. We conclude that threshold-based models with BCS indicators as predictors are useful to understand disease risk (e.g., anestrus), but conversely, they are useless to predict such multicausal disease events at the herd level.

A simplified scoring system for the diagnosis of diarrhea and respiratory diseases in dairy calves.

Objective: To compare the capacity of a simplified calf health scoring chart (SIM score) with the University of Wisconsin's calf health scoring chart (WIN score) for the diagnosis of calf diarrhea and calf respiratory disease (RD). Animals and procedures: Holstein calves (N = 222) were clinically evaluated for diarrhea and RD diagnosis using the WIN and SIM scores. The WIN score was based on fecal consistency for diagnosis of diarrhea (0 = feces of normal consistency to 3 = watery feces; score ≥ 2 = positive diagnosis); and on nasal discharge, ocular discharge, coughing, ear position, and rectal temperature for diagnosis of RD (each clinical sign receives a score of 0 to 3; aggregate score ≥ 5 = positive diagnosis). The SIM score was based on a hide cleanliness score for diagnosis of diarrhea [0 = negative (calf was clean) and 1 = positive (tail head region, thighs, and/or legs were soiled)]; and on nasal discharge, ocular discharge, coughing, and ear position for diagnosis of RD (rectal temperature measurement was not required and each clinical sign had 2 levels of severity; aggregate score ≥ 5 = positive diagnosis). Results: In the RD diagnosis, the SIM score had a sensitivity of 88.24%, a specificity of 95.01%, a positive predictive value (PPV) of 55.56%, and a negative predictive value (NPV) of 99.13%. In the diarrhea diagnosis, the SIM score had a sensitivity of 94.62%, a specificity of 49.64%, a PPV of 18.22%, and an NPV of 98.73%. Conclusion: Compared with the WIN score, the SIM score is a reliable test for diagnosing RD but not for diagnosing diarrhea.

Un système de notation simplifié pour le diagnostic de la diarrhée et des maladies respiratoires chez les veaux laitiers. Objectif: Comparer la capacité d'un tableau de notation simplifié de la santé du veau (score SIM) avec le tableau de notation de la santé du veau (score WIN) de l'University of Wisconsin pour le diagnostic de la diarrhée du veau et de la maladie respiratoire du veau (RD). Animaux et procédures: Des veaux Holstein (N = 222) ont été évalués cliniquement pour le diagnostic de diarrhée et de RD à l'aide des scores WIN et SIM. Le score WIN était basé sur la consistance fécale pour le diagnostic de diarrhée (0 = fèces de consistance normale à 3 = fèces aqueuses; score ≥ 2 = diagnostic positif ); et sur l'écoulement nasal, l'écoulement oculaire, la toux, la position des oreilles et la température rectale pour le diagnostic de RD (chaque signe clinique reçoit un score de 0 à 3; score global ≥ 5 = diagnostic positif ). Le score SIM était basé sur un score de propreté de la peau pour le diagnostic de diarrhée [0 = négatif (le mollet était propre) et 1 = positif (la région de la tête de la queue, les cuisses et/ou les pattes étaient souillées); et sur l'écoulement nasal, l'écoulement oculaire, la toux et la position des oreilles pour le diagnostic de RD (la mesure de la température rectale n'était pas requise et chaque signe clinique avait 2 niveaux de gravité; score global ≥ 5 = diagnostic positif ). Résultats: Dans le diagnostic de RD, le score SIM avait une sensibilité de 88,24 %, une spécificité de 95,01 %, une valeur prédictive positive (VPP) de 55,56 % et une valeur prédictive négative (VPN) de 99,13 %. Dans le diagnostic de diarrhée, le score SIM avait une sensibilité de 94,62 %, une spécificité de 49,64 %, une VPP de 18,22 % et une VPN de 98,73 %. Conclusion: Comparé au score WIN, le score SIM est un test fiable pour diagnostiquer le RD mais pas pour diagnostiquer la diarrhée.(Traduit par Dr Serge Messier).

Association of bovine viral diarrhea virus, bovine herpesvirus 1, and Neospora caninum with late embryonic losses in highly supplemented grazing dairy cows.

The objectives of this study were: 1- to evaluate the association of Bovine Viral Diarrhea Virus (BVDV), Bovine Herpes Virus 1 (BoHV-1), and Neospora caninum (N. caninum) with the risk for Late Embryonic Loss (LEL) in grazing dairy cows, 2- to evaluate blood progesterone concentration at the time of LEL occurrence, and 3- to describe a novel ultrasound-guided technique for conceptus sampling. We run a prospective cohort study involving 92 cows (46 LEL and 46 NLEL). An LEL cow was that having an embryo with no heartbeat, detached membranes, or floating structures, including embryo remnants detected at pregnancy check by ultrasonography (US) 28-42 days post-AI, whereas an NLEL cow was that with embryo heartbeats detectable by US at pregnancy check 28-42 d post-IA. We took two blood samples from every cow at pregnancy check by US (the day of LEL detection) and 28 d later to perform serological diagnosis of BVDV, BoHV-1, and N. caninum; and to measure blood progesterone concentration at pregnancy check (28-42 d post-AI). We also sampled the conceptus from all the LEL cows. We performed PCR to detect BVDV, BoHV-1, and N. caninum in sampled conceptuses from LEL cows. Finally, we evaluated the associations of risk factors (serological titers, seroconversion, and progesterone) with LEL odds with logistic models. The risk for LEL was associated with serological titers to BVDV (P = 0.03) and tended to be associated with seroconversion to BVDV, given that 19.6% (9/46) in LEL and 6.5% (3/46) in NLEL cows seroconverted to BVDV (P = 0.09). In addition, BVDV was detected in conceptuses from LEL cows that seroconverted to BVDV but not in LEL cows that did not seroconvert. Conversely, the risk for LEL was not associated with the titers or seroconversion to BoHV-1 and N. caninum. BoHV-1 and N. caninum were not identified in any of the conceptuses. Finally, blood progesterone concentration was similar in LEL and NLEL cows, and it was not associated with the risk for LEL (P = 0.54). In conclusion, BVDV infection is a risk factor for LEL in dairy cows.

Lameness in Early Lactation Is Associated with Lower Productive and Reproductive Performance in a Herd of Supplemented Grazing Dairy Cows.

The main aim of this study was to assess the associations between the timing of lameness clinical case occurrence in lactation with productive and reproductive performances in grazing Holstein cows. A cohort study was carried out on a dataset with records from a commercial dairy herd (Buenos Aires, Argentina) for cows that calved and were dried off from January 2010 through June 2017. The first recorded event of lameness per lactation was considered for the study. Criteria for lactation inclusion included not having uterine diseases, mastitis, or anovulatory cysts during the studied risk period (i.e., up to 200 DIM). Therefore, a total of 7156 out of 20,086 lactations were included in the statistical analysis. The association between lameness case occurrence in lactation (cows not lame (LG0) vs. lame cows between parturition and first service (LG1) vs. lame cows between first service and first pregnancy (LG2)) with productive (i.e., accumulated milk yield to 150 DIM (MILK150) and 300 DIM (MILK305)) and reproductive performances (hazard of insemination and pregnancy) was analyzed with linear regression models and proportional hazard regression models, respectively. Lame cows produced 161 and 183 kg less MILK150 and MILK305 than non-lame herd mates, respectively. Moreover, LG1 cows produced 216 kg less MILK150 and 200 kg less MILK305 than LG0 cows, and LG2 cows also produced 58 kg less MILK150 and 158 kg less MILK305 than LG0 cows. The LG1 cows had a lower hazard of service than LG0 cows (HR = 0.43, 95%CI = 0.39-0.47). Furthermore, LG1 cows had a lower hazard of pregnancy than LG0 cows (HR = 0.52, 95%CI = 0.46-0.59) and took longer to get pregnant than LG0 cows (median [95%CI], 139 [132-144] vs. 101 [99-103]). Moreover, LG2 cows had a much lower hazard of pregnancy than LG0 cows (HR = 0.08, 95%CI = 0.05-0.12) and much longer calving to first pregnancy interval than LG0 cows (188 [183-196] vs. 101 [99-103]). In conclusion, cows that become lame in early lactation produce less milk and have lower hazards of insemination and pregnancy than herd mates that are healthy or become lame later in lactation. In addition, cows that become lame immediately after the voluntarily waiting period have the poorest reproductive performance (i.e., they have the lowest hazard of pregnancy and the longest calving to pregnancy interval).

Analysis of puerperal metritis treatment records in a grazing dairy farm in Argentina.

OBJECTIVE: To assess the efficacy of antibiotic usage for the treatment of puerperal metritis (PM) and its association with reproductive performance, a retrospective cohort study including a total of 9168 records of cows from a dairy farm in Argentina was run. MATERIAL AND METHODS: Cows having a PM3 (metricheck, scale 0-3) and treated with ceftiofur (ceftiofur crystalline free acid, 6.6 mg/kg) at 0-21 days postpartum (p. p.) (n = 2688), and cows having a PM 1-2 and not treated with an antibiotic at 0-21 days p. p. (n = 6480) were included in the study. All cows were reexamined with metricheck to assess the clinical cure (vaginal discharge [VD] score 0), partial cure (VD score similar or lower than previous), no cure (VD score higher than previous). Cows with a metricheck VD1-3 after 0-21 days p. p. were diagnosed as clinical endometritis (CE) 1-3. The occurrence of PM1-3, cure rate, calving to conception interval, the hazard of pregnancy, odds for non-pregnancy, and odds for CE were analyzed using SAS software. RESULTS: A total of 8876 PM1-3 records were included, 2435 records of PM3 treatments with ceftiofur (27.43 %), and 6441 records of PM1-2 (72.57 %) with no treatment. Cows having PM1 and PM2 became pregnant 14 and 12 days earlier than cows with PM3 (p < 0.001). The PM3 ceftiofur treated cows had a clinical cure of 24.85 % (PM0); 53.63 % had a partially cure; and 18.52 % no cure. Conversely, cows with PM1-2 had a 51.96 %, 20.70 %, and 24.53 % cure rate, respectively (p < 0.001). Cows having complete cure became pregnant 13 and 11 days earlier than cows having partial cure and no cure (p < 0.001). Cows that had PM3 during the first 21 days p. p. had twice the chances of developing CE compared to cows having PM1-2 (41.28 % vs. 24.14 %, p < 0.001). After 21 days p. p., less than 1 % of cows with clinical cure developed CE compared to 63.32 % that developed CE with partial cure, and 38.21 % with no cure (p < 0.001). CONCLUSION AND CLINICAL RELEVANCE: After ceftiofur treatment, 78 % of cows were cured when measured by disappearance of fetid VD but only 25 % of cows had clinical cure when measured by appearance of a clear VD. The cows that remained with clinical metritis had more chances of having CE after 21 days p. p. and had more days open than cows with clear normal VD.

Late embryonic losses in supplemented grazing lactating dairy cows: Risk factors and reproductive performance.

The main objective of this study was to evaluate the risk factors for late embryonic loss (LEL) in supplemented grazing dairy cows. Additional objectives were to assess the incidence of LEL and its association with the reproductive performance of cows. A data set containing productive, reproductive, and health records of 13,551 lactations was used. A retrospective case-control study involving 631 cows with LEL (cases) and 2,524 controls (4 controls per case within each study year) was run. A case of LEL was defined when the embryo had no heartbeat or there was evidence of detached membranes or floating structures including embryo remnants by ultrasonography (US) at 28 to 42 d post-artificial insemination (AI), whereas a non-case was defined as a cow diagnosed with positive pregnancy by US 28 to 42 d post-AI and reconfirmed as pregnant 90 ± 7 d post-AI. Four controls per case were randomly selected from the non-cases with a temporal matching criterion (±3 d around the date of the fecundating AI of the case). Multivariable logistic models were offered with the following predictors: year of LEL (2011 through 2015), season of LEL (summer vs. fall vs. winter vs. spring), parity (1 vs. 2 vs. ≥3), uterine disease (UD), non-uterine disease (NUD), body condition score at parturition, body condition score at 28 to 42 d post-AI (BCS-LEL), days in milk (DIM), and daily milk yield (MY). Statistical significance was set at P < 0.05 and a tendency was set at P ≤ 0.10. We found that 4.7, 22, and 23% of cows had LEL, UD, and NUD, respectively. Cases tended to have higher daily MY than controls (32.5 vs. 31.8 kg); also, cases had much longer calving to pregnancy interval (226 vs. 118 d), lower hazard of pregnancy [hazard ratio = 0.39, 95% confidence interval (CI) = 0.35-0.43], and higher odds for non-pregnancy [odds ratio (OR) = 2.89, 95% CI = 2.37-3.54] than controls. We found that the odds for LEL increased with parity number (OR = 2.48, 95% CI = 1.99-3.08 for parity ≥3) and with BCS-LEL <2.50 (OR = 1.81, 95% CI = 1.33-2.47). Conversely, the odds for LEL decreased with BCS-LEL >3.00 (OR = 0.70, 95% CI = 0.53-0.91). The odds for LEL increased with UD (OR = 1.23, 95% CI = 1.01-1.49), NUD (OR = 1.24, 95% CI = 1.01-1.54), DIM (OR = 1.03, 95% CI = 1.00-1.05), and daily MY (OR = 1.14, 95% CI = 1.04-1.25) in univariable models only. Finally, the odds for LEL were not associated with year, season, DIM, and body condition score at parturition. In conclusion, LEL is associated with extended calving to pregnancy interval, and among its risk factors are parity number and BCS-LEL.

Associations of subclinical hypocalcemia with fertility in a herd of grazing dairy cows.

The main objective was to assess the associations of subclinical hypocalcemia (SCH), diagnosed at parturition (SCH-0) and 7 d in milk (SCH-7), with fertility in a herd of grazing dairy cows. Additional objectives were to characterize Ca concentration on 0 and 7 d in milk (DIM), assessing the risk factors for SCH-0 and SCH-7 and also the relationship with health status (metritis, endometritis, subclinical ketosis, and culling). A prospective observational study was carried out in a dairy farm in Argentina. Holstein cows (n = 126) were body condition scored (BCS, 1-5) on -21 ± 3, 0, 7 ± 3, and 28 ± 7 DIM and blood was collected on 0 and 7 ± 3 DIM to determine Ca and ß-hydroxybutyrate concentrations. Calcium concentrations <2.0 and <2.14 mmol/L were used to define SCH-0 and SCH-7, respectively. The associations of SCH with (1) the odds for pregnancy to first service (P1AI) and pregnancy by 100 DIM (P100) were evaluated by logistic models, (2) the services per pregnancy was evaluated by a Poisson regression model, and (3) the hazards of insemination and pregnancy were evaluated with proportional hazards regression models whereas median days from calving to first insemination and to pregnancy were estimated by Kaplan-Meier method. Additionally, Ca concentration was assessed by linear regression models, and the associations of SCH-0 and SCH-7 with the odds for metritis, endometritis, subclinical ketosis, and culling were evaluated by logistic models. Calcium concentrations were similar at 0 and 7 DIM (2.40 vs. 2.41 mmol/L, respectively); they were higher in cows calving in fall than in summer (2.58 vs. 2.24 mmol/L), and they also were higher in primiparous than in multiparous cows (2.53 vs. 2.28 mmol/L, respectively). The proportion of cows having SCH-0 and SCH-7 was 27.3 and 39.3%, respectively. Fall-calving cows had lower odds for SCH-0 [odds ratio (OR) = 0.31, 95% confidence interval (CI) = 0.12-0.86] than summer-calving cows, multiparous cows had higher odds for SCH-0 (OR = 3.96, 95% CI = 1.09-14.39) than primiparous cows, and cows with prepartum BCS ≥3.00 had higher odds for SCH-0 (OR = 4.03, 95% CI = 1.17-13.89) than in cows with BCS <3.00. Conversely, parity and prepartum BCS were not important predictors for SCH-7. Surprisingly, SCH-0 was not a risk factor for SCH-7. Cows with SCH-0 had lower odds for P1AI (OR = 0.26, 95% CI = 0.07-0.99) than normocalcemic cows, given that P1AI was 14 versus 38%, respectively. The hazard of first service was not associated with SCH-0 (hazard ratio = 1.03, 95% CI = 0.63-1.70) but cows with SCH-0 had lower hazard of pregnancy (hazard ratio = 0.39, 95% CI = 0.16-0.98) and took 32 d longer to get pregnant (105 vs. 73) than normocalcemic cows. Conversely, SCH-7 was not associated with fertility. Finally, SCH-0 and SCH-7 were associated with the odds for subclinical ketosis and metritis, respectively. In conclusion, SCH-0 but not SCH-7 is associated with reduced fertility in a herd of grazing dairy cows, but both were associated with health status.

Identification of Escherichia coli and Trueperella pyogenes isolated from the uterus of dairy cows using routine bacteriological testing and Fourier transform infrared spectroscopy.

BACKGROUND: Uterine disorders are common postpartum diseases in dairy cows. In practice, uterine treatment is often based on systemic or locally applied antimicrobials with no previous identification of pathogens. Accurate on-farm diagnostics are not available, and routine testing is time-consuming and cost intensive. An accurate method that could simplify the identification of uterine pathogenic bacteria and improve pathogen-specific treatments could be an important advance to practitioners. The objective of the present study was to evaluate whether a database built with uterine bacteria from European dairy cows could be used to identify bacteria from Argentinean cows by Fourier transformed infrared (FTIR) spectroscopy. Uterine samples from 64 multiparous dairy cows with different types of vaginal discharge (VD) were collected between 5 and 60 days postpartum, analyzed by routine bacteriological testing methods and then re-evaluated by FTIR spectroscopy (n = 27). RESULTS: FTIR spectroscopy identified Escherichia coli in 12 out of 14 samples and Trueperella pyogenes in 8 out of 10 samples. The agreement between the two methods was good with a Kappa coefficient of 0.73. In addition, the likelihood for bacterial growth of common uterine pathogens such as E. coli and T. pyogenes tended to increase with VD score. The odds for a positive result to E. coli or T. pyogenes was 1.88 times higher in cows with fetid VD than in herdmates with clear normal VD. CONCLUSIONS: We conclude that the presence of E. coli and T. pyogenes in uterine samples from Argentinean dairy cows can be detected with FTIR with the use of a database built with uterine bacteria from European dairy cows. Future studies are needed to determine if FTIR can be used as an alternative to routine bacteriological testing methods.

Effect of peer instruction on the likelihood for choosing the correct response to a physiology question.

The aims of the present study were to measure the effects of individual answer (correct vs. incorrect), individual answer of group members (no vs. some vs. all correct), self-confidence about the responses (low vs. mid vs. high), sex (female vs. male students), and group size (2-4 students) on the odds for change and for correctness after peer instruction in a veterinary physiology course (n = 101 students). Data were assessed by multivariable logistic regression analysis. The likelihood for change after peer instruction increased when the confidence on an individual answer was low (P < 0.01), when the answer was incorrect (P < 0.01), and when group members had different responses (P < 0.01). The likelihood for correctness after peer instruction increased when the confidence in group answers was high (P < 0.01), when the individual answer was correct (P < 0.01), and when at least one of the group members had the correct response (P < 0.01). After peer discussion, more changes were from incorrect to correct responses than vice versa (72% vs. 28%, P < 0.01). Changes to correct answers occurred after discussion with peers having both the correct individual response (76% of times) and also the incorrect individual answer (24% of times). In conclusion, the benefits of peer instruction are due to students having correct answers generally prevail in discussions. Also, students who all have incorrect answers can get the correct answer through debate and discussion.

Dynamics of bacteriologic and cytologic changes in the uterus of postpartum dairy cows.

The objectives of this study were to characterize clinical, intrauterine, bacteriologic and cytologic changes during the first month after parturition in healthy dairy cows and in cows with subclinical endometritis (SE) or clinical endometritis (CE). Furthermore, risk factors related to clinical bacteriologic and cytologic findings were determined. A total of 170 calvings were enrolled, and intrauterine samples were collected on Days 0, 3, 9, 15, 21, and 28 postpartum using the cytobrush technique. The presence of Escherichia coli and Trueperella pyogenes was determined by Fourier transform infrared spectroscopy. The cows were categorized according to their uterine health status (UHS) on Day 21 as healthy (clear or absent vaginal discharge and <5% polymorphonuclear cells [PMN] in the cytologic sample), SE (clear or absent vaginal discharge and ≥5% PMN), or CE (vaginal mucus containing any signs of pus). The prevalence of SE and CE on Day 21 was 27.9% and 58.4%, respectively. Generally, samples from cows with SE and CE showed a greater bacterial growth density (BGD) than those from healthy cows. The BGD tended to be affected by the interaction of time by UHS (P = 0.057). Differences between healthy, SE, and CE cows were found from Day 3 to the last sampling day. Furthermore, the percentage of PMN differed between healthy, SE, and CE cows and was affected by time in a cubic way (decrease/increase/decrease). Overall, E coli was found in 25.4% of the samples, and T pyogenes was identified in 30.2% of the samples. The risk for CE was increased by BGD and the presence of T pyogenes. Conversely, the presence of E coli had no effect on the risk of CE or the risk of SE. The risk for an infection with T pyogenes was greater in the first-parity cows and in cows with assisted calving. In conclusion, changes in BGD and proportion of PMN varied with the UHS (healthy, SE, and CE), which was affected by the presence of T pyogenes but not E coli.

How does a hopping kangaroo breathe?

We developed a model to demonstrate how a hopping kangaroo breathes. Interestingly, a kangaroo uses less energy to breathe while hopping than while standing still. This occurs, in part, because rather than using muscle power to move air into and out of the lungs, air is pulled into (inspiration) and pushed out of (expiration) the lungs as the abdominal organs "flop" within the kangaroo's body. Specifically, as the kangaroo hops upward, the abdominal organs lag behind, and the insertion of the diaphragm is pulled toward its origin, flattening the dome and increasing the vertical dimension of the thoracic cavity (the thoracic cavity and lungs enlarge). Increasing the volume of the thoracic cavity reduces alveolar pressure below atmospheric pressure (barometric pressure), and air moves into the alveoli by bulk flow. In contrast, the impact of the organs against the diaphragm at each landing causes expiration. Specifically, upon landing, the abdominal organs flop into the diaphragm, causing it to return to its dome shape and decreasing the vertical dimension of the thoracic cavity. This compresses the alveolar gas volume and elevates alveolar pressure above barometric pressure, so air is expelled. To demonstrate this phenomenon, the plunger of a syringe model of the respiratory system was inserted through a compression spring. Holding the syringe and pressing the plunger firmly against a hard surface expels air from the lungs (the balloon within the syringe deflates) and compresses the spring. This models the kangaroo landing after a hop forward. Subsequently, the compression spring provides the energy for the "kangaroo" to "hop" forward upon the release of the syringe, and air enters the lungs (the balloon within the syringe inflates). The model accurately reflects how a hopping kangaroo breathes. A model was chosen to demonstrate this phenomenon because models engage and inspire students as well as significantly enhance student understanding.

Hooke's law: applications of a recurring principle.

Students generally approach topics in physiology as a series of unrelated phenomena that share few underlying principles. However, if students recognized that the same underlying principles can be used to explain many physiological phenomena, they may gain a more unified understanding of physiological systems. To address this concern, we developed a simple, inexpensive, and easy to build model to demonstrate the underlying principles regarding Starling's Law of the Heart as well as lung and arterial elastic recoil. A model was chosen because models significantly enhance student understanding. Working with models also encourages research-oriented learning and helps our students understand complex ideas. Students are drawn into discussion by the power of learning that is associated with manipulating and thinking about objects. Recognizing that the same underlying principles can be used to explain many physiological phenomena may help students gain a more complete understanding of physiological systems.

A model of locomotor-respiratory coupling in quadrupeds.

Locomotion and respiration are not independent phenomena in running mammals because locomotion and respiration both rely on cyclic movements of the ribs, sternum, and associated musculature. Thus, constraints are imposed on locomotor and respiratory function by virtue of their linkage. Specifically, locomotion imposes mechanical constraints on breathing that require the respiratory cycle to be synchronized with gait. Thus, many mammals, including humans, synchronize respiration with the movement of the limbs during locomotion. For example, quadrupeds synchronize locomotor and respiratory cycles at a 1:1 ratio (stride/breath) over a wide range of speeds. Interestingly, quadrupeds maintain an almost constant stride frequency (and therefore respiratory frequency) at different speeds. To increase speed, quadrupeds lengthen their stride. Accordingly, to increase minute ventilation, quadrupeds must increase tidal volume since respiratory rate is coupled with stride frequency. We developed a simple, inexpensive, and easy to build model to demonstrate this concept. A model was chosen because models significantly enhance student understanding. Students are drawn into discussion by the power of learning that is associated with manipulating and thinking about objects. Building and using this model strengthen the concept that locomotor-respiratory coupling provides a basis for the appropriate matching of lung ventilation to running speed and metabolic power.

Student interaction characteristics during collaborative group testing.

We used collaborative testing in a veterinary physiology course (65 students) to answer the following questions: 1) do students with individual correct responses or students with individual incorrect responses change their answers during group testing? and 2) do high-performing students make the decisions, that is, are low-performing students carried by high-performing peers? To address these questions, students first completed the exam in the traditional format as individuals. After completing the exam as individuals, students completed the same exam in groups of two. Finally, the same questions were discussed by the instructor and students (instructor feedback). We found that students with individual incorrect responses changed their answers during group testing more often than students with individual correct responses (odds ratio: 7.58, P < 0.01). Furthermore, student feedback was more beneficial when group members had different individual answers than when they had same individual answers (P < 0.05). In addition, when group members had different individual answers, more answers were changed to correct responses than to incorrect responses (77% vs. 23%, P < 0.01). It was more important to have the correct answer than to be the high-performing student, because the student with the correct response (being either the high- or low-performing student) generally prevailed ( approximately 80% of the time, P = 0.5). Finally, the positive effects of group testing (77% of total effects, P < 0.05) were due to students who changed their individual answer to the correct response after discussion with peers with the correct response and also with the incorrect individual response.

Collaborative group testing benefits high- and low-performing students.

We used collaborative group testing in a veterinary physiology course (65 students) to test the hypothesis that all students (e.g., high-performing and low-performing students of each group) benefit from collaborative group testing. In this format, students answered questions in the traditional format as individuals. Immediately after completing the exam as individuals, students answered the same questions in groups of two, and, finally, the same questions were discussed by the instructor and students. We measured two learning outcomes for every student: individual and group test scores. Based on individual test scores, students were categorized as "high performing" (students with higher individual scores) or "low performing" (students with lower individual scores). Finally, student evaluations of the format were collected. Collaborative group testing enhanced student performance. Specifically, group scores were higher than individual scores (P < 0.001). Importantly, the size of the collaborative testing effect was large for the population and for the low-performing students; however, the collaborative testing effect was small for the high-performing students. Finally, student evaluations of this testing format were very positive. In conclusion, collaborative group testing was beneficial for all students; however, collaborative testing was significantly more beneficial for low-performing students.