Evaporative water loss from dairy cows in climate-controlled respiration chambers.

The effects of ambient temperature (AT) on total evaporative water loss from dairy cows at different relative humidity (RH) and air velocity (AV) levels were studied. Twenty Holstein dairy cows with an average parity of 2.0 ± 0.7 and body weight of 687 ± 46 kg participated in the study. Two climate-controlled respiration chambers were used. The experimental indoor climate was programmed to follow a diurnal pattern with AT at night being 9°C lower than during the day. Night AT was gradually increased from 7 to 21°C and day AT was increased from 16°C to 30°C within an 8-d period, both with an incremental change of 2°C/d. The effect of 3 RH levels with a diurnal pattern were studied as well, with low values during the day and high values during the night: low (day, 30%; night, 50%), medium (day, 45%; night, 70%), and high (day, 60%; night, 90%). The effects of AV were studied during the daytime at 3 levels: no fan (0.1 m/s), fan at medium speed (1.0 m/s), and fan at high speed (1.5 m/s). The medium and high AV levels were only combined with medium RH. In total, there were 5 treatments with 4 replicates each. The animals had free access to feed and water. Based on the water balance principle inside the respiration chambers, the total evaporative water loss from dairy cows at a daily level was quantified by measuring the mass of water in the incoming and outgoing air, condensed water, added water from a humidifier, and evaporative water from a wet floor, drinking bowl, manure reservoir, and water bucket. Water evaporation from a sample skin area was measured with a ventilated skin box, and water evaporation, through respiration with a face mask. The results show that RH/AV levels had no significant effect on total evaporative water loss, whereas the interaction effect between RH/AV with AT was significant. Cows at a high RH had a tendency for a lower increasing rate of evaporative water loss compared with cows at a low RH (0.61 vs. 0.79 kg/d per 1°C increase of AT). Cows at medium and high AV levels had a greater increasing rate than cows at low AV (0.91 and 0.95 vs. 0.71 kg/d per 1°C increase of AT, respectively). The increase of evaporative heat loss from dairy cows was mainly a result of the increase in evaporation (of sweat) from the skin. The skin water evaporation determined with the water balance method (less evaporation from respiration) and the ventilated skin box method showed no significant difference. The implication of this study is that cows at a high AT depend mainly on evaporative cooling from the skin. The ventilated skin box method, measuring only a small part of the skin during a short period during the day, can be a convenient and accurate way to determine the total cutaneous evaporative water loss from cows.

Effects of increasing air temperature on skin and respiration heat loss from dairy cows at different relative humidity and air velocity levels.

The focus of this study was to identify the effects of increasing ambient temperature (T) at different relative humidity (RH) and air velocity (AV) levels on heat loss from the skin surface and through respiration of dairy cows. Twenty Holstein dairy cows with an average parity of 2.0 ± 0.7 and body weight of 687 ± 46 kg participated in the study. Two climate-controlled respiration chambers were used. The experimental indoor climate was programmed to follow a diurnal pattern with ambient T at night being 9°C lower than during the day. Night ambient T was gradually increased from 7 to 21°C and day ambient T was increased from 16 to 30°C within an 8-d period, both with an incremental change of 2°C per day. A diurnal pattern for RH was created as well, with low values during the day and high values during the night (low: RH_l = 30-50%; medium: RH_m = 45-70%; and high: RH_h = 60-90%). The effects of AV were studied during daytime at 3 levels (no fan: AV_l = 0.1 m/s; fan at medium speed: AV_m = 1.0 m/s; and fan at high speed: AV_h = 1.5 m/s). The AV_m and AV_h were combined only with RH_m. In total, there were 5 treatments with 4 replicates (cows) for each. Effects of short and long exposure time to warm condition were evaluated by collecting data 2 times a day, in the morning (short: 1-h exposure time) and afternoon (long: 8-h exposure time). The cows were allowed to adapt to the experimental conditions during 3 d before the main 8-d experimental period. The cows had free access to feed and water. Sensible heat loss (SHL) and latent heat loss (LHL) from the skin surface were measured using a ventilated skin box placed on the belly of the cow. These heat losses from respiration were measured with a face mask covering the cow's nose and mouth. The results showed that skin SHL decreased with increasing ambient T and the decreasing rate was not affected by RH or AV. The average skin SHL, however, was higher under medium and high AV levels, whereas it was similar under different RH levels. The skin LHL increased with increasing ambient T. There was no effect of RH on the increasing rate of LHL with ambient T. A larger increasing rate of skin LHL with ambient T was observed at high AV level compared with the other levels. Both RH and AV had no significant effects on respiration SHL or LHL. The cows lost more skin sensible heat and total respiration heat under long exposure than short exposure. When ambient T was below 20°C the total LHL (skin + respiration) represented approx. 50% of total heat loss, whereas above 28°C the LHL accounted for more than 70% of the total heat loss. Respiration heat loss increased by 34 and 24% under short and long exposures when ambient T rose from 16 to 32°C.

Effects of increasing air temperature on physiological and productive responses of dairy cows at different relative humidity and air velocity levels.

This study determined the effects of increasing ambient temperature (T) at different relative humidity (RH) and air velocity (AV) levels on the physiological and productive responses of dairy cows. Twenty Holstein dairy cows were housed inside climate-controlled respiration chambers, in which the climate was programmed to follow a daily pattern of lower night and higher day T with a 9°C difference, excluding effects from sun radiation. Within our 8-d data collection period, T was gradually increased from 7 to 21°C during the night (12 h) and 16 to 30°C during the day (12 h), with an incremental change of 2°C per day for both nighttime and daytime T. During each research period, RH and AV were kept constant at 1 of 5 treatment levels. A diurnal pattern for RH was created, with lower levels during the day and higher levels during the night: low (RH_l: 30-50%), medium (RH_m: 45-70%), and high (RH_h: 60-90%). The effects of AV were studied during the day at 3 levels: no fan (AV_l: 0.1 m/s), fan at medium speed (AV_m: 1.0 m/s), and fan at high speed (AV_h: 1.5 m/s). Effects of short and long exposure time to increasing T were evaluated by collecting data 2 times a day: in the morning [short: 1 h (or less) - exposure time] and afternoon (long: 8 h - exposure time). The animals had free access to feed and water and both were ad libitum. Respiration rate (RR), rectal temperature (RT), skin temperature (ST), dry matter intake, water intake, milk yield, and composition were measured. The inflection point temperatures (IPt) at which a certain variable started to change were determined for the different RH and AV levels and different exposure times. Results showed that IPt under long exposure time for RR (first indicator) varied between 18.9 and 25.5°C but was between 20.1 and 25.9°C for RT (a delayed indicator). The IPt for both RR and RT decreased with higher RH levels, whereas IPt increased with higher AV for RR but gave a minor change for RT. The ST was positively correlated with ambient T and ST was not affected by RH but significantly affected by AV. For RR, all IPt was lower under long exposure time than under short exposure time. The combination of higher RH levels and low AV level negatively affected dry matter intake. Water intake increased under all treatments except RH_l-AV_l. Treatment RH_h-AV_l negatively affected milk protein and fat yield, whereas treatments RH_m-AV_m and RH_m-AV_h reduced milk fat yield. We concluded that RH and AV significantly affected the responses of RR, RT, ST, and productive performance of high-producing Holstein cows. These responses already occurred at moderate ambient T of 19 to 26°C.

A genotyping array for the globally invasive vector mosquito, Aedes albopictus.

BACKGROUND: Although whole-genome sequencing (WGS) is the preferred genotyping method for most genomic analyses, limitations are often experienced when studying genomes characterized by a high percentage of repetitive elements, high linkage, and recombination deserts. The Asian tiger mosquito (Aedes albopictus), for example, has a genome comprising up to 72% repetitive elements, and therefore we set out to develop a single-nucleotide polymorphism (SNP) chip to be more cost-effective. Aedes albopictus is an invasive species originating from Southeast Asia that has recently spread around the world and is a vector for many human diseases. Developing an accessible genotyping platform is essential in advancing biological control methods and understanding the population dynamics of this pest species, with significant implications for public health. METHODS: We designed a SNP chip for Ae. albopictus (Aealbo chip) based on approximately 2.7 million SNPs identified using WGS data from 819 worldwide samples. We validated the chip using laboratory single-pair crosses, comparing technical replicates, and comparing genotypes of samples genotyped by WGS and the SNP chip. We then used the chip for a population genomic analysis of 237 samples from 28 sites in the native range to evaluate its usefulness in describing patterns of genomic variation and tracing the origins of invasions. RESULTS: Probes on the Aealbo chip targeted 175,396 SNPs in coding and non-coding regions across all three chromosomes, with a density of 102 SNPs per 1 Mb window, and at least one SNP in each of the 17,461 protein-coding genes. Overall, 70% of the probes captured the genetic variation. Segregation analysis found that 98% of the SNPs followed expectations of single-copy Mendelian genes. Comparisons with WGS indicated that sites with genotype disagreements were mostly heterozygotes at loci with WGS read depth < 20, while there was near complete agreement with WGS read depths > 20, indicating that the chip more accurately detects heterozygotes than low-coverage WGS. Sample sizes did not affect the accuracy of the SNP chip genotype calls. Ancestry analyses identified four to five genetic clusters in the native range with various levels of admixture. CONCLUSIONS: The Aealbo chip is highly accurate, is concordant with genotypes from WGS with high sequence coverage, and may be more accurate than low-coverage WGS.

Associations between patient-reported late effects and systemic cytokines in long-term survivors of head and neck cancer treated with radiotherapy.

PURPOSE: Head and neck cancer (HNC) treatment may lead to late effects and impaired health-related quality of life of survivors. Knowledge on long-term late effects after radiotherapy (RT) and potential underlying biological mechanisms is lacking. We assessed the prevalence of xerostomia, dysphagia, and chronic fatigue (CF) in HNC survivors ≥ 5 years post-RT, and examined associations between pro-inflammatory cytokines and late effects. METHODS: In a cross-sectional study, 263 HNC survivors treated between 2007 and 2013 were enrolled. They completed validated questionnaires assessing xerostomia and dysphagia (the EORTC QLQ-H&N35), and CF (the Fatigue Questionnaire), and underwent blood sampling and clinical examination. Pro-inflammatory cytokines were analyzed in 262 survivors and 100 healthy age- and gender-matched controls. RESULTS: Median time since treatment was 8.5 years. The proportions of survivors reporting xerostomia, dysphagia, and CF were 58%, 31%, and 33%, respectively, with a preponderance of females. We found no significant associations between IL-6, IL-8, IP-10, TARC, TNF, or ENA-78 and the three late effects. The odds of having elevated levels of IL-6 and IP-10 were significantly higher in the survivors compared to the controls. CONCLUSIONS: More than one-third of long-term HNC survivors experienced xerostomia, dysphagia, and CF. Persistent inflammation, with elevated systemic cytokines, was not associated with these late effects, although HNC survivors had higher levels of some cytokines than the controls. IMPLICATIONS FOR CANCER SURVIVORS: This study provides new knowledge on late effects that can serve as grounds for informing patients with HNC about risk of late effects more than 5 years after RT.

Effects of serum-free storage on morphology, phenotype, and viability of ex vivo cultured human conjunctival epithelium.

The use of amniotic membrane (AM) represents one of the major developments in ocular surface reconstruction. However, in a study on patients with primary pterygium, transplantation of AM with ex vivo expanded human conjunctival epithelial cells (HCjE) promoted earlier epithelialization than AM alone. We previously showed that cultured human limbal epithelial cells maintain their morphology, phenotype, and viability for one week when stored at 23°C. The current study investigates the feasibility of storing HCjE in HEPES-MEM and Optisol-GS at 23°C for 4 and 7 days, respectively. The five experimental groups were analyzed by light microscopy, immunohistochemistry, transmission electron microscopy, and a viability assay. The ultrastructural integrity of cultured HCjE was well preserved following 4 days of storage, however, 7 days of storage resulted in some loss of cell-cell contacts and epithelial detachment from the amniotic membrane. The number of microvilli in cultured HCjE not subjected to storage was 2.03±0.38 microvilli/µm. In comparison, after 4 and 7 days of HEPES-MEM storage this number was 1.69±0.54 microvilli/µm; P=0.98 and 0.89±1.0 microvilli/µm; P=0.28, respectively. After Optisol-GS storage for 4 and 7 days, the mean number of microvilli was 1.07±1.0 microvilli/µm; P=0.47 and 0.07±0.07 microvilli/µm; P=0.03, respectively. The number of cell layers in cultured HCjE not subjected to storage was 4.4±0.3 cell layers, as opposed to 4.0±0.9 cell layers; P=0.89 after 4 days of HEPES-MEM storage and 2.8±0.6 cell layers; P=0.01 after 7 days of storage in HEPES-MEM. The number of cell layers after 4 and 7 days of storage in Optisol-GS was 3.7±0.2 cell layers; P=0.46 and 3.4±0.4 cell layers; P=0.18, respectively. The expression of markers for undifferentiated cells (ΔNp63α, ABCG2 and p63), proliferating cells (Ki67 and PCNA), goblet cells (Ck7 and MUC5AC), stratified squamous epithelial cells (Ck4), and apoptotic cells (caspase-3) in cultured HCjE appeared to be unchanged after 4 and 7 days of HEPES-MEM and Optisol-GS storage. The percentage of viable cells in cultured HCjE not subjected to storage (91.4%±3.2%) was sustained after 4 and 7 days of storage in HEPES-MEM (94.1%±4.5%; P=0.99 and 85.1%±13.7%; P=0.87, respectively) as well as after 4 and 7 days of storage in Optisol-GS (87.7%±15.2%; P=0.97 and 79.8%±15.7%; P=0.48, respectively). We conclude that cultured HCjE may be stored for at least 4 days in serum-free conditions at 23°C while maintaining the phenotype and viability. HEPES-MEM appears to be comparable to Optisol-GS for serum-free storage with preservation of the ultrastructure for at least 4 days.

Genetic screening for von Hippel-Lindau gene mutations in non-syndromic pheochromocytoma: low prevalence and false-positives or misdiagnosis indicate a need for caution.

Genetic testing of tumor susceptibility genes is now recommended in most patients with pheochromocytoma or paraganglioma (PPGL), even in the absence of a syndromic presentation. Once a mutation is diagnosed there is rarely follow-up validation to assess the possibility of misdiagnosis. This study prospectively examined the prevalence of von Hippel-Lindau (VHL) gene mutations among 182 patients with non-syndromic PPGLs. Follow-up in positive cases included comparisons of biochemical and tumor gene expression data in 64 established VHL patients, with confirmatory genetic testing in cases with an atypical presentation. VHL mutations were detected by certified laboratory testing in 3 of the 182 patients with non-syndromic PPGLs. Two of the 3 had an unusual presentation of diffuse peritoneal metastases and substantial increases in plasma metanephrine, the metabolite of epinephrine. Tumor gene expression profiles in these 2 patients also differed markedly from those associated with established VHL syndrome. One patient was diagnosed with a partial deletion by Southern blot analysis and the other with a splice site mutation. Quantitative polymerase chain reaction, multiplex ligation-dependent probe amplification, and comparative genomic hybridization failed to confirm the partial deletion indicated by certified laboratory testing. Analysis of tumor DNA in the other patient with a splice site alteration indicated no loss of heterozygosity or second hit point mutation. In conclusion, VHL germline mutations represent a minor cause of non-syndromic PPGLs and misdiagnoses can occur. Caution should therefore be exercised in interpreting positive genetic test results as the cause of disease in patients with non-syndromic PPGLs.

Phytoextraction of heavy metals by willows growing in biosolids under field conditions.

Biosolids produced by sewage treatment facilities can exceed guideline thresholds for contaminant elements. Phytoextraction is one technique with the potential to reduce these elements allowing reuse of the biosolids as a soil amendment. In this field trial, cuttings of seven species/cultivars of Salix(willows) were planted directly into soil and into biosolids to identify their suitability for decontaminating biosolids. Trees were irrigated and harvested each year for three consecutive years. Harvested biomass was weighed and analyzed for the contaminant elements: As, Cd, Cu, Cr, Hg, Pb, Ni, and Zn. All Salix cultivars, except S. chilensis, growing in soils produced 10 to 20 t ha(-1) of biomass, whereas most Salix cultivars growing in biosolids produced significantly less biomass (<6 t ha(-1)). Salix matsudana (30 t ha(-1)) and S. × reichardtii A. Kerner (18 t ha(-1)) had similar aboveground biomass production in both soil and biosolids. These were also the most successful cultivars in extracting metals from biosolids, driven by superior biomass increases and not high tissue concentrations. The willows were effectual in extracting the most soluble/exchangeable metals (Cd, 0.18; Ni, 0.40; and Zn, 11.66 kg ha(-1)), whereas Cr and Cu were extracted to a lesser degree (0.02 and 0.11 kg ha(-1)). Low bioavailable elements, As, Hg, and Pb, were not detectable in any of the aboveground biomass of the willows.

Mosquitoes and their potential predators in rice agroecosystems of the Mekong Delta, southern Vietnam.

Culex tritaeniorhynchus, Cx. gelidus, and Cx. quinquefasciatus, known vectors of Japanese encephalitis (JE), are distributed in rice agroecosystems in Asian countries. Very few integrated studies on the breeding habitats of rice-field mosquitoes, including JE vectors, have been conducted in Vietnam. We investigated the mosquito fauna and potential predators in 8 rice growing areas in the Mekong Delta region of southern Vietnam, during the wet and dry seasons of 2009. Mosquitoes and their predators were collected from a variety of aquatic habitats (rice fields, ponds, wetlands, shrimp ponds, ditches, canals, and rivers). We collected 936 Culex spp. (354 Cx. tritaeniorhynchus, 240 Cx. vishnui s.l., 189 Cx. fuscocephala, and 42 Cx. gelidus), 33 Uranotaenia, 25 Anopheles, and 9 Mimomyia (4 Mi. chamberlaini) in the dry season. During the rainy season, we collected 1,232 Culex spp. (132 Cx. vishnui s.l., 66 Cx. tritaeniorhynchus, 9 Cx. gelidus, 4 Cx. fuscocephala, and 2 Cx. bitaeniorhynchus), 236 Anopheles spp. (40 An. vagus and 1 An. sinensis), and 7 Uranotaenia (3 Ur. lateralis). Heteroptera such as Micronecta, Veliidae, and Pleidae were abundant and widely distributed in both seasons. Based on a stepwise generalized linear model, the abundance of mosquitoes and their predators in rice fields was high when the rice plant length was short and water depth was shallow. Therefore, the use of insecticides during the earlier stages of rice growth should be avoided in order to preserve the predator populations.