Low colostrum yield in Jersey cattle and potential risk factors.

Consumption of an adequate volume of high-quality colostrum is vital to a dairy calf's ability to survive and become a productive herd member. However, some dairy herds have reported a deficiency of colostrum production, which ranges from a low volume to no colostrum produced, by cows during fall and winter. Little information regarding this phenomenon exists. The purpose of this study was to characterize the syndrome and identify potential risk factors for low colostrum yield. A 2,500-cow Jersey dairy farm was enrolled in a prospective cohort study in May 2016, to evaluate possible effects of photoperiod, temperature, and cow factors on colostrum production. Dairy personnel were trained to collect, weigh, and evaluate colostrum quality. Information on parity, previous lactation length, previous 305-d mature-equivalent milk production, and dry period length were collected through the farm's dairy management software. Weather and photoperiod data were also collected. Over the year of enrollment, 2,988 eligible cows calved and had colostrum weights recorded and 38% were primiparous (n = 1,143), 25% were in their second lactation (n = 752), and 37% were in their third or greater lactation (n = 1,093). The overall average colostrum yield was 6.6 kg/cow in June 2016, 2.5 kg/cow in December 2016, and 4.8 kg/cow in May 2017. Multiparous cows had a larger decline in colostrum production between June and December (6.6 to 1.3 kg/cow) compared with primiparous animals (6.5 to 4.2 kg/cow). Overall, average colostrum production decreased by 0.17 kg/cow per week during this time, 0.22 kg for multiparous cows and 0.08 kg for primiparous cows. A logistic regression model was constructed for all cows to evaluate effects of cow factors on low colostrum production (<2.7 kg at first milking). Dry period length, calf sex, singleton or twin, age at freshening, month of calving and previous lactation length were significantly associated with the probability of low colostrum yield (<2.7 kg at first milking). A cross-correlation function analysis between the time series for colostrum yield and photoperiod revealed a high correlation at the time of calving and 1 mo prior, particularly for multiparous cows. A pedigree analysis showed that extreme colostrum yield (low vs. high) followed some sire lines. Low colostrum production in this herd could have an economic effect on the dairy and calf health and appears to have a strong seasonal and, potentially, a genetic component.

Association of abnormal hind-limb postures and back arch with gait abnormality in dairy cattle.

Detection of lameness in individual cows is important for the prompt treatment of this painful and production-limiting disease. Current methods for lameness detection involve watching cows walk for several strides. If clinical signs predictive of lameness could be observed more conveniently, as cows are undergoing regularly scheduled examinations while standing, detection levels could increase. The objective of this study was to assess the association between postures observed while cows are standing in stanchions and clinical lameness evaluated by locomotion scoring, and to evaluate the observation of these postures as a test for lameness. The study included 1,243 cows from 4 farms. Cows were observed while standing in stanchions for regularly scheduled management procedures and the presence of arched back and cow-hocked, wide-stance, and favored-limb postures were recorded. The same cows were locomotion-scored as they exited the milking parlor. The proportion of cows observed with arched back and cow-hocked and favored-limb postures increased with increasing severity of lameness (higher locomotion score) but did not increase for the wide-stance posture. For the presence of these postures as a test for lameness (locomotion score ≥3), sensitivity and specificity were 0.63 and 0.64 for back arch, 0.54 and 0.57 for cow hocks, and 0.05 and 0.98 for favored limb. Back-arched, cow-hocked, and favored limb postures were associated with lameness but were not highly sensitive or specific as diagnostic tests. However, observation of back arch may be useful to identify cows needing further examination.

Comparison of modeled sampling strategies for estimation of dairy herd lameness prevalence and cow-level variables associated with lameness.

Monitoring herd lameness prevalence has utility for dairy producers and veterinarians in their efforts to reduce lameness, for animal welfare assessment programs, and for researchers. Locomotion scoring is a method used to quantify lameness and calculate prevalence. Because of the time necessary to locomotion score each cow in large dairy herds, a sampling strategy to determine herd lameness prevalence that allows scoring of fewer cows would be useful. Such a sampling strategy must be validated for accuracy compared with the lameness prevalence when all cows in a herd are locomotion scored. The purpose of this study was to assess 3 previously suggested methods of estimating lameness prevalence by strategic sampling of dairy herds. Sampling strategies tested included (1) sampling a calculated number of cows in the middle third of the milking parlor exit order for each pen, (2) sampling a calculated number of cows weighted across pens and distributed evenly within each pen, and (3) sampling all cows in the high production, low production, and hospital pens. Lactating cows on 5 dairy farms in Washington and Oregon (n=4,422) were locomotion scored using a 5-point scale to determine herd-level lameness prevalence (percentage with locomotion score ≥3). Milking parlor exit order, order in headlocks at the feed bunk within each pen, and breed were recorded for each cow. The number of days in lactation, milk production, and parity were collected from farm computer records. Pen grouping strategy for each farm was obtained by interview with farm management. Sampling strategies were modeled using the locomotion score data set for each herd. Estimates of lameness prevalence obtained from the milking parlor exit order sample and the sample distributed across pens were within 5 percentage points of the whole herd prevalence. The third strategy estimated the lameness prevalence within 5 percentage points on 4 farms, but overestimated prevalence on 1 farm. Pen-level prevalence obtained by locomotion score of all cows in the pen was variable and not reliably predictive of herd-level prevalence. Cows of Holstein breed, parity >1, and exiting the milking parlor in the last 20% of the pen had greater odds of lameness compared with other breeds, parities, and milking parlor exit order groups in a multivariate analysis. This study indicates that the sampling strategies using the middle of milking parlor exit order and a calculated sample distributed across the herd may be used to obtain an estimate of herd lameness prevalence.

Retrospective evaluation of health event data recording on 50 dairies using Dairy Comp 305.

The purpose of this study was to characterize health data recording in herds using Dairy Comp 305 (Valley Agricultural Software, Tulare, CA), focused on the most common diseases of dairy cattle: mastitis, uterine infection (metritis), and diseases causing lameness. Herds using Dairy Comp 305 were chosen for the study because it was the most commonly used program in the United States (35% of operations using computer records/60% of cows on operations using computer records), the authors were familiar with the program, and there was convenient access to herds using the program. Specific objectives were to (1) determine the percentage of herds recording mastitis, metritis, and diseases causing lameness and the number of user-defined events used to record those diseases, (2) identify the information recorded in the remarks about the event used for each disease, and (3) evaluate the consistency of health event remarks recorded. A convenience sample of 50 Dairy Comp 305 compressed cow files was obtained directly from dairies that the authors had contact with or files obtained from industry consultants. The 50 herds included in the study were from 9 different states: California (n=3), Colorado (n=2), Iowa (n=2), Idaho (n=8), Minnesota (n=2), New Mexico (n=9), Oregon (n=2), Texas (n=2), Utah (n=1), and Washington (n=19). The average number of milking cows of the herds was 3,053 (median=2,217), ranging from 310 to 12,490 cows. The majority of dairies in this study were recording health events associated with mastitis, metritis, and diseases causing lameness. However, as reported previously, most health records observed in the current study lacked the accuracy and consistency needed to be useful for evaluating and informing herd-level health management decisions. This situation likely reflects the intended use of those records by farm personnel and the user-defined nature of health records in the absence of accepted industry standards or recommendations for health data-recording practices.

Short communication: Effects of dairy calf hutch elevation on heat reduction, carbon dioxide concentration, air circulation, and respiratory rates.

Heat stress affects dairy calf welfare and can result in morbidity, mortality, and lower weight gain. The purpose of this project was to evaluate the effects of elevating the back of plastic calf hutches on measures of ventilation and heat stress. A total of 15 calves housed in individual hutches were enrolled, with each calf hutch serving as its own control. Heat, humidity, carbon dioxide, and wind speed were measured inside each hutch and the observations were compared with external measurements over two 24-h periods; 1 period without and 1 with hutch elevation. Respiratory rates were measured in the morning and afternoon as an indicator of the degree of heat stress experienced by calves with and without elevation of the hutch. When the hutch was elevated, internal hutch temperatures were cooler than external temperatures, hutch carbon dioxide levels were lower and respiratory rates were lower, particularly comparing the afternoon observation periods.

Presynchronization with GnRH 7 days prior to resynchronization with CO-Synch did not improve pregnancy rate in lactating dairy cows.

The objective was to determine the effect of presynchronization with GnRH 7 d prior to the initiation of resynchronization with CO-Synch on pregnancy/AI (P/AI) of resynchronization in lactating dairy cows, and the effect of GnRH on P/AI from previous breeding. All parity Holstein cows (n = 3287) from four dairy farms were enrolled. Cows not detected in estrus by 28 ± 3 d (Day -7) after a previous breeding were assigned to receive either GnRH (100 µg, im; n = 1636) or no GnRH (Control; n = 1651). Cows not detected in estrus during the 7 d after GnRH underwent pregnancy diagnosis (35 ± 3 d after previous breeding, Day 0); non-pregnant cows (n = 1232) in the Control (n = 645) and GnRH (n = 587) groups were resynchronized with a CO-Synch protocol. Briefly, cows received 100 µg GnRH on Day 0, 25 mg PGF(2α) on Day 7, and 72 h later (Day 10) were given 100 µg GnRH and concurrently inseminated. Serum progesterone concentrations (n = 55 cows) were elevated in 47.3, 70.9, and 74.5% of cows on Days -7, 0, and 7, respectively. The proportion of cows with high progesterone concentrations on Day -7 and Day 0 were 44.1% and 88.2% (P < 0.003), and 55.2% and 33.2% (P > 0.1), for GnRH and Control groups, respectively. Accounting for significant variables such as locations (P < 0.0001) and parity categories (P < 0.05), the P/AI (35 ± 3 d after AI) for resynchronization was not different between GnRH and Control groups [26.7% (95% CI: 23.2, 30.5; (157/587) vs 28.4% (95% CI: 25.0, 31.9; (183/645); P > 0.1]. There were no significant location by treatment or parity by treatment interactions. Accounting for significant variables such as location (P < 0.0001) and parity categories (P < 0.001), the P/AI was not different between GnRH and Control groups for the previous service [60.2%; 95% CI: 57.9, 62.6; (986/1636) vs 59.1%; 95% CI: 56.7, 61.5; (976/1651); P > 0.1)]. There were no significant location by treatment or parity by treatment interactions. In conclusion, more cows presynchronized with GnRH 7 d prior to resynchronization with CO-Synch had elevated progesterone concentrations at initiation of resynchronization than those not presynchronized. The GnRH treatment 7 d prior to resynchronization with CO-Synch, when given 28 ± 3 d after a previous breeding, did not improve P/AI in lactating dairy cows; furthermore, compared to the control, it did not significantly affect pregnancy rate from the previous breeding.

Factors associated with the rectal temperature of Holstein dairy cows during the first 10 days in milk.

Daily evaluation of rectal temperature (RT) during the first 10 d in milk (DIM) is used to facilitate the early identification of postpartum complications, particularly metritis in dairy cows. The factors associated with RT of postpartum dairy cows have not been clearly established and the RT threshold used to define fever has been variable. The objectives were to identify factors associated with the RT of postpartum dairy cows and provide descriptive statistics of the RT during the first 10 DIM to clarify the normal range of RT for cows. Daily RT was evaluated from 1 to 10 DIM for all cows calving during 2 consecutive summers on a single 1,500-cow Holstein dairy. Cows were placed into metabolic/digestive (METB), infectious (INF), and no recorded disease (NONE) groups based on disease diagnoses during the first 10 DIM. Cows were grouped based on calving difficulty and parity. Multiple linear regression models with repeated measures were used to evaluate the factors associated with RT. Three hundred and ninety-two cows were evaluated, of which 45% were primiparous and 32% required assistance at calving. No difference was observed in calving assistance by parity. First disease diagnoses peaked in the INF and METB groups at 3 and 1 DIM, respectively. The RT of primiparous cows was 0.1 to 0.2°C higher than that of multiparous cows from 1 to 8 DIM, accounting for calving difficulty, twin births, month of calving, and disease group in the model. The INF group cows had a higher RT than did NONE group cows (38.9±0.04 to 39.2±0.73 vs. 38.7±0.03°C, respectively) on each of the first 10 DIM, which was approximately 0.6°C higher from 3 to 5 DIM. The RT of cows with metritis was at least 0.1°C higher (38.8±0.05°C) than that of NONE group cows beginning 4 d before diagnosis. The mean RT of primiparous, defined healthy (NONE group) cows was 38.8±0.02°C, with an upper normal limit (mean+2 SD) of 39.6°C. The mean RT of multiparous cows in the NONE group during the first 10 DIM was 38.7±0.01°C, with an upper normal limit of 39.5°C. The RT of dairy cattle during the first 10 DIM was associated with parity, month of calving, and an infectious disease diagnosis, particularly the diagnosis of metritis. The normal RT of dairy cattle in the immediate postpartum period, during the warm summer months, is potentially higher than that generally reported.

Incidence and transmission of Mycoplasma bovis mastitis in Holstein dairy cows in a hospital pen: A case study.

The objective was to determine the incidence and transmission of mycoplasma mastitis in the hospital pen in a dairy herd of 650 lactating cows after a hospital pen was established following an outbreak of this disease. Mycoplasma mastitis status was monitored for 3 months through repeated collection of milk samples from cows with clinical mastitis (CM) and from bulk tank milk. During the outbreak 13 cows were diagnosed with Mycoplasma bovis CM, 1 cow with Mycoplasma sp. mastitis and 8 cows showed signs of arthritis, 3 of which were confirmed as having M. bovis arthritis. M. bovis isolates from cows with CM, arthritis and bulk tank milk had indistinguishable chromosomal digest pattern fingerprints. Incidence rates of M. bovis CM cases in the milking and hospital pens were 0.01 and 1.7 cases per 100 cow-days at risk. Approximately 70% of cows with M. bovis CM became infected within 12 days of entering the hospital pen. Transmission of M. bovis in the hospital pen occurred as 3 episodes. Each episode corresponded to the introduction of a cow with M. bovis CM from a milking pen. Evidence indicates that cows with M. bovis CM from milking pens were the source of transmission of the disease in the hospital pen and thus their presence in the hospital pen appeared to be a risk factor for transmission of M. bovis mastitis in this single case study herd.

Factors associated with concentrations of select cytokine and acute phase proteins in dairy cows with naturally occurring clinical mastitis.

The objective of the current observational study was to determine the potential associations between cow factors, clinical mastitis (CM) etiology, and concentrations of select acute phase proteins and cytokines in milk from affected quarters of cows with CM. Cows with CM (n=197) were grouped based on systemic disease severity, milk culture result, parity, days in milk (DIM), previous CM occurrence, and season of the year when CM occurred. Concentrations of lipopolysaccharide-binding protein (LBP), haptoglobin (Hp), BSA, IFN-gamma, tumor necrosis factor-alpha (TNF-alpha), IL-1beta, IL-8, IL-10, IL-12, transforming growth factor (TGF)-alpha, and TGF-beta and activity of lactate dehydrogenase (LDH) were evaluated. Differences in the least squares means log(10) transformed concentrations of these proteins were compared using multiple linear regression mixed models. The milk concentrations of LBP, Hp, IL-1beta, IL-10, and IL-12, and activity of LDH in milk were higher in cows with moderate to severe versus mild systemic disease. The concentrations of Hp, BSA, IL-1beta, and IL-10 in milk were higher in cows with a gram-negative versus gram-positive milk culture result. Season of the year when CM occurred was associated with the concentration of all proteins evaluated except for IL-1beta and IL-12. Concentrations were higher in the winter versus summer except for Hp and TGF-beta, for which the opposite was true. Concentrations of LBP, IL-10, and IL-12, and LDH activity in milk were associated with DIM group. Except for LBP, these proteins were lower in cows with CM during the first 60 DIM versus those in mid or later lactation. Interferon-gamma, TNF-alpha, and IL-8 were undetectable in 67, 31, and 20% of samples, respectively. Detection of IFN-gamma and IL-8 was associated with season, and detection of TNF-alpha and IL-8 was associated with systemic disease severity. The current study provides the most comprehensive report of milk concentrations of innate immune response proteins in cows with naturally occurring CM and identifies factors that potentially influence those concentrations. Further investigation into the seasonal variation of cytokine production and its potential effect on the outcome of CM is warranted. Furthermore, the results of this study provide useful data for planning future studies examining the role of the innate immune response in CM.

Herd management practices and their association with bulk tank somatic cell count on United States dairy operations.

The objective of this study was to evaluate associations between bulk tank somatic cell count (BTSCC) and herd management practices using data collected in the National Animal Health Monitoring System Dairy 2002 study. Twenty-six percent and 17.8% of 1,013 operations reported a BTSCC < 200,000 cells/mL and > 400,000 cells/mL, respectively. Univariate analysis identified associations between management variables and BTSCC. The use of mattresses, sand, and newspaper as bedding were all associated with a lower BTSCC. Primary lactating cow housing facility, outside maternity housing area, flooring type cows walk or stand on, and use of automatic take-offs were also associated with BTSCC. Multivariate associations between management variables and BTSCC were determined by backward elimination ordinal logistic regression. The odds of an operation from the West, Midwest, and Northeast having a high BTSCC were lower than those from the Southeast. The odds of a higher BTSCC were 2 times greater for operations with a rolling herd average milk production < 9,090 kg/cow per year compared with those with > or = 9,090 kg/cow per year. Operations using composted manure were 2.9 times more likely to have a higher BTSCC than those not using composted manure. Finally, operations that reported not using a coliform mastitis vaccine were 1.7 times more likely to have a higher BTSCC than those using one. Future studies of the association between management practices and BTSCC should include an evaluation of the quality of management practice application and herd prevalence of contagious mastitis pathogens. Significant variables identified in this study dealt with housing, use of composted manure for bedding, and coliform mastitis vaccine use, suggesting the effect of environmental mastitis pathogens may be more influential on BTSCC than previously thought.

Escherichia coli isolates' serotypes, genotypes, and virulence genes and clinical coliform mastitis severity.

Dairy cattle with clinical mastitis caused by Escherichia coli exhibit a wide range of disease severity, from mild, with only local inflammatory changes of the mammary gland, to severe, with significant systemic derangement. The present study was designed to examine the relationship between serotype and virulence genes of E. coli mastitis isolates, different levels of systemic disease severity, and farm from which the E. coli strain was obtained. One hundred twenty-three E. coli milk isolates were obtained from cows with clinical mastitis of varying systemic disease severity from 6 different farms. No predominant serotype was identified by farm or by systemic disease severity; however, the most frequent serotype, O158:NM (n = 3), was isolated from cows in the moderate severity group. Virulence genes evaluated were identified infrequently and were not associated with systemic disease severity. Evaluation of genetic similarity showed no clustering assigned by farm or mastitis severity based on systemic disease signs. We concluded that a high degree of genotypic variability is characteristic of E. coli strains causing clinical mastitis within and between different farms and systemic severity groups, and that specific cow factors probably play a more important role in determining systemic disease severity.

Short communication: Efficacy of parenteral ceftiofur for treatment of systemically mild clinical mastitis in dairy cattle.

The objective of this study was to evaluate the effect of intramuscular (i.m.) ceftiofur (2.2 mg/kg) on important outcomes of systemically mild clinical mastitis episodes in lactating dairy cattle. Cows with clinical mastitis were randomly assigned to a treatment group: pirlimycin intramammary (i.m.m.) (n = 35), pirlimycin i.m.m. and ceftiofur i.m.m. (n = 36), cephapirin i.m.m. (n = 40), cephapirin i.m. and ceftiofur i.m. (n = 33). Sixty-nine, 22, and 9% of initial cultures were gram-negative, gram-positive, and mixed, respectively. Logistic regression analysis showed no significant associations between treatment groups and loss of quarter, recurrence, or culling. Mixed infections, positive milk culture at 7 d after leaving hospital pen, decreased rumen motility, and absence of udder firmness were associated with increased odds of mastitis recurrence. The results suggest that i.m. ceftiofur treatment has no beneficial effects on the outcome of systemically mild clinical mastitis.

Bacteremia associated with naturally occuring acute coliform mastitis in dairy cows.

OBJECTIVE: To determine the incidence of bacteremia in dairy cows with naturally occurring acute coliform mastitis (ACM) with a wide range of disease severity. DESIGN: Cohort study. ANIMALS: 144 dairy cows with ACM from 6 herds. PROCEDURE: Cows were examined at time of identification of ACM (time 0) and classified as having mild, moderate, or severe mastitis on the basis of rectal temperature, hydration status, rumen contraction rate, and attitude. Cows were reexamined at 24 or 48 hours. Bacteriologic culturing of milk and blood (30 ml), CBC, and serum biochemical analysis were performed at each time point. Appropriate samples were obtained at a single point from herdmates without mastitis (controls) that were closely matched for lactation number and days since parturition. Blood culture results were compared among severity groups and controls by use of chi2 tests, as was outcome of an ACM episode for cows grouped by blood bacterial isolates. RESULTS: Bacteria were isolated from 52 blood samples from 46 of 144 (32%) cows with ACM, which was significantly more than control cows (11/156; 7.1%). Group-1 isolates (Escherichia coli, Pasteurella multocida, Mannheimia haemolytica, Klebsiella pneumoniae, Enterobacter agglomerans, and Salmonella enterica serotype Typhimurium) were identified in 20 of 144 (14%) cows with ACM and 0 of 156 control cows. Group-1 isolates were identified in 4.3, 9.1, and 42% of cows classified as having mild, moderate, and severe ACM, respectively. Escherichia coli and K pneumoniae milk and blood isolates obtained from the same cow were of the same genotype. Bacillus spp were identified in 21 of 144 (15%) cows with ACM, which was significantly more than control cows (3/156; 1.9%). Thirty-five percent of cows with a group-1 isolate died during the mastitis episode. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that bacteremia develops in a substantial proportion of cows with ACM. Classification of severity of disease is important for establishment of effective treatment protocols; parenteral antimicrobial treatment may be indicated in cows with ACM.

Use of systemic disease signs to assess disease severity in dairy cows with acute coliform mastitis.

OBJECTIVE: To evaluate the use of systemic disease signs for classifying severity of acute coliform mastitis in dairy cows. DESIGN: Prospective cohort study. ANIMALS: 144 dairy cows. PROCEDURE: Cows were examined at the time of initial identification of disease (time 0) and classified as having mild, moderate, or severe disease on the basis of rectal temperature, hydration status, rumen contraction rate, and attitude. A CBC and serum biochemical analyses were performed, and milk samples were submitted for bacterial culture at time 0 and 48 hours later. RESULTS: 69 cows were classified as having mild disease, 44 as having moderate disease, and 31 as having severe disease. Median WBC and neutrophil counts were significantly lower in cows with moderate or severe disease at time 0 than in cows with mild disease. Band neutrophil count was significantly higher at 48 hours and serum calcium concentration was significantly lower at time 0 and at 48 hours in cows with severe or moderate disease, compared with cows with mild disease. Twenty-eight, 51, and 77% of cows with mild, moderate, and severe disease, respectively, had > 100,000 colony-forming units/ml of milk at time 0. The odds that a cow with severe disease would die or be culled were 3.6 times the odds for a cow with moderate disease and 11.2 times the odds for a cow with mild disease. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a classification scheme based on readily observable systemic disease signs can be used to classify disease severity in cows with acute coliform mastitis.

Characterization of stage-specific tyrosinated alpha-tubulin immunoperoxidase staining patterns in Sertoli cells of rat seminiferous tubules by light microscopic image analysis.

Microtubules are involved in many structural and functional changes that occur in Sertoli cells during the cycle of the seminiferous epithelium. However, few studies have addressed stage-specific changes in the distribution of microtubules that accompany the process of spermatogenesis. This study provides a stage-by-stage immunohistochemical evaluation of Sertoli cell microtubules in paraffin sections of whole rat testes using an antibody to tyrosinated alpha-tubulin. Microtubules that contain tyrosinated tubulin are considered to be less stable and are therefore expected to populate Sertoli cells undergoing dynamic changes during spermatogenesis. A quantitative method was developed to analyze the relative tyrosinated alpha-tubulin staining in different stages of the cycle. Immunostaining patterns of the stages were separated into five different groups. Stages VII-VIII had the least amount of tyrosinated alpha-tubulin, while stages IX-XI contained the greatest amount. The staining patterns were consistent with established structural changes in the seminiferous epithelium, such as the formation of ectoplasmic specializations, the presence of microtubule nucleation sites along the periphery of the apical cytoplasm, and the translocation of elongate spermatids from deep crypts in the Sertoli cell to the tubule lumen. These data should provide improved methods for the evaluation of microtubules in the study of Sertoli cell responses to environmental toxicants and testicular diseases.

Baculovirus gp64 envelope glycoprotein is sufficient to mediate pH-dependent membrane fusion.

The baculovirus gp64 envelope glycoprotein is a major component of the envelope of the budded virus (BV) and is involved in BV entry into the host cell by endocytosis. To determine whether gp64 alone was sufficient to mediate membrane fusion, the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus gp64 protein was transiently expressed in uninfected insect cells. Cells expressing the baculovirus gp64 protein were examined for membrane fusion activity by using a syncytium formation assay under various conditions of exposure to low pH. Cells expressing the gp64 protein mediated membrane fusion and syncytium formation in a pH-dependent manner. A pH of 5.5 or lower was required to induce membrane fusion. In addition, exposure of gp64-expressing cells to low pH for as little as 5 s was sufficient to induce gp64-mediated syncytium formation. These studies provide direct evidence that gp64 is a pH-dependent membrane fusion protein and suggest that gp64 is the protein responsible for fusion of the virion envelope with the endosome membrane during BV entry into the host cell by endocytosis.

Unilateral renal agenesis in chick embryos: a model for chronic renal insufficiency.

Although renal agenesis and dysgenesis are relatively common and significant birth defects, no animal model to date has been utilized to adequately study these developmental pathologies. Blockage of the migration of the mesonephric duct in Day 2 chick embryos results in unilateral renal agenesis (URA) on the operated side, thus providing a model of chronic renal insufficiency. Embryos with URA respond with an increase in the rate of growth of the remaining meso- and metanephric kidney. The allometric scaling of single (left) kidney weight to total body weight in control embryos is KM = 3.48M0.98 compared to KM = 3.02M1.16 in embryos with URA. In addition, embryos with URA exhibit a progressively polycystic mesonephros with distinct glomerulonephritis and expansion of the renal tubules. These renal changes are insufficient for normal urine (allantoic fluid) production and oliguria persists throughout incubation. While mortality is unaffected by URA in embryos up to Day 14 of incubation, there is a steady increase in mortality after Day 14; no chick embryo with URA lives beyond Day 18 of the 21-day incubation period.