Spillover of highly pathogenic avian influenza H5N1 virus to dairy cattle.

Highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b virus has caused the death of millions of domestic birds and thousands of wild birds in the U.S. since January, 20221-4 Throughout this outbreak, spillovers to mammals have been frequently documented5-12. We report spillover of HPAI H5N1 virus in dairy cattle herds across several states in the U.S. The affected cows displayed clinical signs encompassing decreased feed intake, altered fecal consistency, respiratory distress, and decreased milk production with abnormal milk. Infectious virus and viral RNA were consistently detected in milk from affected cows. Viral distribution in tissues via immunohistochemistry and in situ hybridization revealed a distinct tropism of the virus for the epithelial cells lining the alveoli of the mammary gland in cows. Whole viral genome sequences recovered from dairy cows, birds, domestic cats, and a raccoon from affected farms indicated multidirectional interspecies transmissions. Epidemiologic and genomic data revealed efficient cow-to-cow transmission after apparently healthy cows from an affected farm were transported to a premise in a different state. These results demonstrate the transmission of HPAI H5N1 clade 2.3.4.4b virus at a non-traditional interface underscoring the ability of the virus to cross species barriers.

Highly Pathogenic Avian Influenza A(H5N1) Clade 2.3.4.4b Virus Infection in Domestic Dairy Cattle and Cats, United States, 2024.

We report highly pathogenic avian influenza A(H5N1) virus in dairy cattle and cats in Kansas and Texas, United States, which reflects the continued spread of clade 2.3.4.4b viruses that entered the country in late 2021. Infected cattle experienced nonspecific illness, reduced feed intake and rumination, and an abrupt drop in milk production, but fatal systemic influenza infection developed in domestic cats fed raw (unpasteurized) colostrum and milk from affected cows. Cow-to-cow transmission appears to have occurred because infections were observed in cattle on Michigan, Idaho, and Ohio farms where avian influenza virus-infected cows were transported. Although the US Food and Drug Administration has indicated the commercial milk supply remains safe, the detection of influenza virus in unpasteurized bovine milk is a concern because of potential cross-species transmission. Continued surveillance of highly pathogenic avian influenza viruses in domestic production animals is needed to prevent cross-species and mammal-to-mammal transmission.

Superhydrophobic 3D-Assembled Metallic Nanoparticles for Trace Chemical Enrichment in SERS Sensing.

The performance of surface-enhanced Raman spectroscopy (SERS) is determined by the interaction between highly diluted analytes and boosted localized electromagnetic fields in nanovolumes. Although superhydrophobic surfaces are developed for analyte enrichment, i.e., to concentrate and transfer analytes toward a specific position, it is still challenging to realize reproducible, uniform, and sensitive superhydrophobic SERS substrates over large scales, representing a major barrier for practical sensing applications. To overcome this challenge, a superhydrophobic SERS chip that combines 3D-assembled gold nanoparticles on nanoporous substrates is proposed, for a strong localized field, with superhydrophobic surface treatment for analyte enrichment. Intriguingly, by concentrating droplets in the volume of 40 µL, the sensitivity of 1 nm is demonstrated using 1,2-bis(4-pyridyl)-ethylene molecules. In addition, this unique chip demonstrates a relative standard deviation (RSD) of 2.2% in chip-to-chip reproducibility for detection of fentanyl at 1 µg mL-1 concentration, revealing its potential for quantitative sensing of chemicals and drugs. Furthermore, the trace analysis of fentanyl and fentanyl-heroin mixture in human saliva is realized after a simple pretreatment process. This superhydrophobic chip paves the way toward on-site and real-time drug sensing to tackle many societal issues like drug abuse and the opioid crisis.

Use of nCounter mRNA profiling to identify at-arrival gene expression patterns for predicting bovine respiratory disease in beef cattle.

BACKGROUND: Transcriptomics has identified at-arrival differentially expressed genes associated with bovine respiratory disease (BRD) development; however, their use as prediction molecules necessitates further evaluation. Therefore, we aimed to selectively analyze and corroborate at-arrival mRNA expression from multiple independent populations of beef cattle. In a nested case-control study, we evaluated the expression of 56 mRNA molecules from at-arrival blood samples of 234 cattle across seven populations via NanoString nCounter gene expression profiling. Analysis of mRNA was performed with nSolver Advanced Analysis software (p < 0.05), comparing cattle groups based on the diagnosis of clinical BRD within 28 days of facility arrival (n = 115 Healthy; n = 119 BRD); BRD was further stratified for severity based on frequency of treatment and/or mortality (Treated_1, n = 89; Treated_2+, n = 30). Gene expression homogeneity of variance, receiver operator characteristic (ROC) curve, and decision tree analyses were performed between severity cohorts. RESULTS: Increased expression of mRNAs involved in specialized pro-resolving mediator synthesis (ALOX15, HPGD), leukocyte differentiation (LOC100297044, GCSAML, KLF17), and antimicrobial peptide production (CATHL3, GZMB, LTF) were identified in Healthy cattle. BRD cattle possessed increased expression of CFB, and mRNA related to granulocytic processes (DSG1, LRG1, MCF2L) and type-I interferon activity (HERC6, IFI6, ISG15, MX1). Healthy and Treated_1 cattle were similar in terms of gene expression, while Treated_2+ cattle were the most distinct. ROC cutoffs were used to generate an at-arrival treatment decision tree, which classified 90% of Treated_2+ individuals. CONCLUSIONS: Increased expression of complement factor B, pro-inflammatory, and type I interferon-associated mRNA hallmark the at-arrival expression patterns of cattle that develop severe clinical BRD. Here, we corroborate at-arrival mRNA markers identified in previous transcriptome studies and generate a prediction model to be evaluated in future studies. Further research is necessary to evaluate these expression patterns in a prospective manner.

Large-Scale, Ion-Current-Based Proteomic Investigation of the Rat Striatal Proteome in a Model of Short- and Long-Term Cocaine Withdrawal.

Given the tremendous detriments of cocaine dependence, effective diagnosis and patient stratification are critical for successful intervention yet difficult to achieve due to the largely unknown molecular mechanisms involved. To obtain new insights into cocaine dependence and withdrawal, we employed a reproducible, reliable, and large-scale proteomics approach to investigate the striatal proteomes of rats (n = 40, 10 per group) subjected to chronic cocaine exposure, followed by either short- (WD1) or long- (WD22) term withdrawal. By implementing a surfactant-aided precipitation/on-pellet digestion procedure, a reproducible and sensitive nanoLC-Orbitrap MS analysis, and an optimized ion-current-based MS1 quantification pipeline, >2000 nonredundant proteins were quantified confidently without missing data in any replicate. Although cocaine was cleared from the body, 129/37 altered proteins were observed in WD1/WD22 that are implicated in several biological processes related closely to drug-induced neuroplasticity. Although many of these changes recapitulate the findings from independent studies reported over the last two decades, some novel insights were obtained and further validated by immunoassays. For example, significantly elevated striatal protein kinase C activity persisted over the 22 day cocaine withdrawal. Cofilin-1 activity was up-regulated in WD1 and down-regulated in WD22. These discoveries suggest potentially distinct structural plasticity after short- and long-term cocaine withdrawal. In addition, this study provides compelling evidence that blood vessel narrowing, a long-known effect of cocaine use, occurred after long-term but not short-term withdrawal. In summary, this work developed a well-optimized paradigm for ion-current-based quantitative proteomics in brain tissues and obtained novel insights into molecular alterations in the striatum following cocaine exposure and withdrawal.

Estimating IgG concentration directly by radial immunodiffusion or indirectly by refractometry measure of serum total protein lack precision.

OBJECTIVE: To establish and compare the precision of serum total protein (STP) measured by an optical refractometer to the precision of IgG concentrations measured using radial immunodiffusion (RID), the reference test for quantifying IgG in neonatal calves. SAMPLE: 6 sera with previously measured IgG concentration using RID from neonatal beef calves were selected from 3 stratum: low-serum IgG stratum between >5.0 and <15.0g/L(n = 4); moderate-serum IgG stratum between 35.0-45.0g/L(n = 1); high-serum IgG stratum between 60.0-70.0g/L(n = 1). METHODS: STP was measured 13 times with an optical refractometer. IgG concentrations were measured 28 times with a commercial bovine IgG RID for each sera. The homogeneity of variance within the tests was evaluated with the Levene test (α = 0.10). Unrestricted random sampling bootstrapping (5,000 repetitions) was used to calculate the coefficient of variation (CV) for each serum and test. The homogeneity of variance between simulated test CVs by serum was evaluated (α = 0.10). Differences between simulated test CV by serum were assessed with the Kruskal-Wallis test (α = 0.05). RESULTS: No difference was observed in the variance for STP between sera (P = .39). The average CV for STP was 4.2%, 10.1% for the low IgG stratum, and 15.5% for the moderate/high IgG stratum. Variance differed in serum IgG concentration (P < .0001). Serum with higher IgG concentrations had more variance. Simulated CV for STP and IgG had homogeneity of variance for only 1 sera (P = .31). STP had a smaller CV compared to IgG for every serum (P < .0001). CLINICAL RELEVANCE: Estimating IgG concentration directly by RID or indirectly by STP lacks the precision that might affect diagnostic interpretation regarding a calf's absorption of maternal antibodies.

Sources of variance in the results of a commercial bovine immunoglobulin G radial immunodiffusion assay.

Radial immunodiffusion (RID) is used to quantify IgG concentration in neonatal beef or dairy calf serum; variability has been noted that may affect the precision and accuracy of assay results. We determined the source, range, and homogeneity of variance in the results of a commercial bovine IgG RID assay (Triple J Farm). To estimate the variance in the precipitin ring diameter, we used 6 sera, measured 28 times across 8 plates and 4 lots, and 3 standards with known IgG concentrations, measured 75 times across 69 plates and 5 lots. The source of diameter variance was determined using variance partition coefficients for lot, plate, and repetition. We used 11 different methods to generate standard curves to convert RID precipitin ring diameters to IgG concentrations. The Levene test of homogeneity of variance (α = 0.1) was used to evaluate the equality of variance between the standards or serum precipitin ring diameters and calculated IgG concentrations. Lot and plate contributed minimally to the diameter variance. Precipitin ring diameters had equal variance. Calculated IgG concentrations for serum not requiring dilution had equal variance. A linear equation from aggregated standards, performed within the same day, had greater accuracy for the calculated IgG concentrations of the standards compared to other equation methods. Regardless of standard curve methodology or IgG concentration, variability inherent to the assay limits its clinical usefulness.

Placentophagia and the Tao of POEF.

Placentophagia, ingestion of placenta and amniotic fluid, usually during parturition, is a behavioral feature of nearly all nonaquatic, placental mammals, and is a nexus for several interlocking behavioral phenomena. Placentophagia has not been typical of human cultures, but in recent years, some women in affluent societies have engaged in it, thereby bringing publicity to the behavior. First, we summarized benefits of placentophagia for nonhuman mammals, which include increased attractiveness of neonates, enhanced onset of maternal behavior, suppression of pseudopregnancy, and enhancement of opioid hypoalgesia by Placental Opioid-Enhancing Factor (POEF), a benefit that may extend well outside the context of parturition. The research on POEF in animals was discussed in detail. Then we discussed placentophagia (placentophagy) in humans, and whether there is validity to the claims of various benefits reported primarily in the pro-placentophagy literature, and, although human afterbirth shows POEF activity, the POEF effect has not yet been tested in humans. Finally, we discussed the general possible implications, for the management of pain and addiction, of isolating and characterizing POEF.

Assessment of accredited veterinary diagnostic laboratory use of breakpoints for canine and feline Escherichia coli infections in the United States and Canada.

The objective of this study was to assess the use of breakpoints in antibiotic susceptibility testing among veterinary diagnostic laboratories in the United States and Canada. An eight-question survey was conducted via phone and email to determine how often laboratories use breakpoints consistent with published guidelines in wounds, lower urinary tract infections and upper urinary tract infections (pyelonephritis) involving Escherichia coli, both in dogs and cats, for a total of 6 different hypothetical clinical scenarios. Nineteen veterinary diagnostic laboratories that perform antibiotic susceptibility testing on samples from dogs and cats in the United States or Canada and were accredited by the American Association of Veterinary Laboratory Diagnosticians (AAVLD) responded to the survey between January 15th and September 15th, 2022. The overall response rate of laboratories that were not excluded for known lack of dog and cat antibiotic susceptibility testing was 19 of 44 laboratories. Of the 17 respondent laboratories that reported using minimal inhibitory concentration breakpoints, only four laboratories used breakpoints consistent with published guidelines in all six clinical scenarios included in the survey. Our results suggest that there is clinically important variation in what breakpoints laboratories use to determine antibiotic susceptibility, which is of antibiotic stewardship and clinical relevance. Using breakpoints that are too high, too low, or inappropriately reporting "not interpreted" as the interpretive category may result in inappropriate use of antibiotics.

Placentophagia in humans and nonhuman mammals: causes and consequences.

Afterbirth ingestion by nonhuman mammalian mothers has a number of benefits: (1) increasing the interaction between the mother and infant; (2) potentiating pregnancy-mediated analgesia in the delivering mother; (3) potentiating maternal brain opioid circuits that facilitate the onset of caretaking behavior; and (4) suppressing postpartum pseudopregnancy. Childbirth is fraught with additional problems for which there are no practical nonhuman animal models: postpartum depression, failure to bond, hostility toward infants. Ingested afterbirth may contain components that ameliorate these problems, but the issue has not been tested empirically. The results of such studies, if positive, will be medically relevant. If negative, speculations and recommendations will persist, as it is not possible to prove the negative. A more challenging anthropological question is "why don't humans engage in placentophagia as a biological imperative?" Is it possible that there is more adaptive advantage in not doing so?

Assessment of Diversity of Antimicrobial Resistance Phenotypes and Genotypes of Mannheimia haemolytica Isolates From Bovine Nasopharyngeal Swabs.

The threat of bovine respiratory disease (BRD) for cattle operations is exacerbated by increasing prevalence of antimicrobial resistance (AMR) in Mannheimia haemolytica, a leading cause of BRD. Characterization of AMR in M. haemolytica by culture and susceptibility testing is complicated by uncertainty regarding the number of colonies that must be selected to accurately characterize AMR phenotypes (antibiograms) and genotypes in a culture. The study objective was to assess phenotypic and genotypic diversity of M. haemolytica isolates on nasopharyngeal swabs (NPS) from 28 cattle at risk for BRD or with BRD. NPS were swabbed onto five consecutive blood agar plates; after incubation up to 20 M. haemolytica colonies were selected per plate (up to 100 colonies per NPS). Phenotype was determined by measuring minimum inhibitory concentrations (MIC) for 11 antimicrobials and classifying isolates as resistant or not. Genotype was indirectly determined by matrix-assisted laser desorption/ionization time of flight mass spectroscopy (MALDI-TOF MS). NPS from 11 of 28 cattle yielded at least one M. haemolytica isolate; median (range) of isolates per NPS was 48 (1-94). NPS from seven cattle yielded one phenotype, 3 NPS yielded two, and 1 NPS yielded three; however, within a sample all phenotypic differences were due to only one MIC dilution. On each NPS all M. haemolytica isolated were the same genotype; genotype 1 was isolated from three NPS and genotype two was isolated from eight. Diversity of M. haemolytica on bovine NPS was limited, suggesting that selection of few colonies might adequately identify relevant phenotypes and genotypes.

Primary Hypoparathyroidism and Recurring Hypocalcemia in a Quarter Horse Gelding-A Case Report.

Hypoparathyroidism is an uncommon endocrine disorder in the horse characterized by a transient or permanent parathyroid hormone insufficiency. Hypoparathyroidism is associated with hypocalcemia and hyperphosphatemia, primarily presenting with clinical signs consistent with hypocalcemia. This case report describes clinical presentation and treatment of a horse with severe hypocalcemia due to primary hypoparathyroidism. A 17-year-old, 542 kg Quarter Horse gelding presented for shaking and tremors. Significant findings include generalized muscle fasciculations, synchronous diaphragmatic flutter, and a markedly hypermetric hindlimb gait. Hematology revealed a moderate hyperkalemia, hyperphosphatemia, hypomagnesemia, and severe hypocalcemia. Initial treatment consisted of oral and intravenous calcium supplementation and fluid therapy. Thirty-six hours after presentation, clinical signs resolved, and treatment was discontinued. Clinical signs reoccurred after the discontinuation of treatment. A presumptive diagnosis of primary hypoparathyroidism was made based on low parathyroid hormone in the presence of low ionized calcium. The patient was maintained on oral calcium carbonate (feed grade lime) and vitamin AED supplementation. Hypoparathyroidism is rare but oral supplementation of calcium with calcium carbonate resulted in a favorable outcome with no apparent decrease in performance. Long-term supplementation may be required to prevent disease recurrence.

Sedation with dexmedetomidine is associated with transient gallbladder wall thickening and peritoneal effusion in some dogs undergoing abdominal ultrasonography.

BACKGROUND: Dexmedetomidine often is used for sedation before or during abdominal ultrasonography. The effect of dexmedetomidine on gallbladder wall thickness is unknown. HYPOTHESIS/OBJECTIVES: To investigate the relationship between dexmedetomidine administration and gallbladder wall thickening in dogs. The hypothesis was that sedation with dexmedetomidine will cause transient gallbladder wall thickening. Gallbladder wall thickness will be associated with duration of sedation and recumbency position. ANIMALS: Seventy-nine client owned dogs and 10 healthy research dogs. METHODS: A prospective observational study (n = 79) was used to establish the prevalence of gallbladder wall thickening (> 2.0 mm) after sedation with dexmedetomidine. A randomized, crossover study (n = 10) was used to evaluate the effect of time and recumbency position on the development of gallbladder wall thickening. Linear mixed models were used. RESULTS: The proportion of client-owned dogs that developed gallbladder wall thickening was 24.05% (19/79; 95% confidence interval [CI], 15.1%-35.0%) with a median dose of dexmedetomidine of 5.0 µg/kg (range, 2.0-12.5 µg/kg). After sedation, the proportion of research dogs that developed gallbladder wall thickening in left lateral (5/10, 50%; 95% CI, 18.7%-81.3%) and dorsal (7/10, 70%; 95% CI, 34.8%-93.3%) recumbency did not differ significantly (P = .45). Gallbladder wall thickening developed within 20 to 40 minutes. Duration of sedation was significantly associated with thickening of the gallbladder wall (P < .001). Five dogs developed 9 instances of peritoneal effusion in both lateral (5) and dorsal (4) recumbency. CONCLUSIONS AND CLINICAL IMPORTANCE: Sedation with dexmedetomidine is associated with gallbladder wall thickening (> 2.0 mm) and peritoneal effusion that could be confused with pathologic etiologies.

Streamlined Multimodal DESI and MALDI Mass Spectrometry Imaging on a Singular Dual-Source FT-ICR Mass Spectrometer.

The study of biological specimens by mass spectrometry imaging (MSI) has had a profound influence in the various forms of spatial-omics over the past two decades including applications for the identification of clinical biomarker analysis; the metabolic fingerprinting of disease states; treatment with therapeutics; and the profiling of lipids, peptides and proteins. No singular approach is able to globally map all biomolecular classes simultaneously. This led to the development of many complementary multimodal imaging approaches to solve analytical problems: fusing multiple ionization techniques, imaging microscopy or spectroscopy, or local extractions into robust multimodal imaging methods. However, each fusion typically requires the melding of analytical information from multiple commercial platforms, and the tandem utilization of multiple commercial or third-party software platforms-even in some cases requiring computer coding. Herein, we report the use of matrix-assisted laser desorption/ionization (MALDI) in tandem with desorption electrospray ionization (DESI) imaging in the positive ion mode on a singular commercial orthogonal dual-source Fourier transform ion cyclotron resonance (FT-ICR) instrument for the complementary detection of multiple analyte classes by MSI from tissue. The DESI source was 3D printed and the commercial Bruker Daltonics software suite was used to generate mass spectrometry images in tandem with the commercial MALDI source. This approach allows for the generation of multiple modes of mass spectrometry images without the need for third-party software and a customizable platform for ambient ionization imaging. Highlighted is the streamlined workflow needed to obtain phospholipid profiles, as well as increased depth of coverage of both annotated phospholipid, cardiolipin, and ganglioside species from rat brain with both high spatial and mass resolution.

Evaluation of buprenorphine in a postoperative pain model in rats.

We evaluated the commonly prescribed analgesic buprenorphine in a postoperative pain model in rats, assessing acute postoperative pain relief, rebound hyperalgesia, and the long-term effects of postoperative opioid treatment on subsequent opioid exposure. Rats received surgery (paw incision under isoflurane anesthesia), sham surgery (anesthesia only), or neither and were treated postoperatively with 1 of several doses of subcutaneous buprenorphine. Pain sensitivity to noxious and nonnoxious mechanical stimuli at the site of injury (primary pain) was assessed at 1, 4, 24, and 72 h after surgery. Pain sensitivity at a site distal to the injury (secondary pain) was assessed at 24 and 72 h after surgery. Rats were tested for their sensitivity to the analgesic and locomotor effects of morphine 9 to 10 d after surgery. Buprenorphine at 0.05 mg/kg SC was determined to be the most effective; this dose induced isoalgesia during the acute postoperative period and the longest period of pain relief, and it did not induce long-term changes in opioid sensitivity in 2 functional measures of the opioid system. A lower dose of buprenorphine (0.01 mg/kg SC) did not meet the criterion for isoalgesia, and a higher dose (0.1 mg/kg SC) was less effective in pain relief at later recovery periods and induced a long-lasting opioid tolerance, indicating greater neural adaptations. These results support the use of 0.05 mg/kg SC buprenorphine as the upper dose limit for effective treatment of postoperative pain in rats and suggest that higher doses produce long-term effects on opioid sensitivity.

Amniotic-fluid ingestion enhances central δ-opioid-induced hypoalgesia in rats in the cold-water tail-flick assay in a repeated-measures design.

Placental Opioid Enhancing Factor (POEF) is found in amniotic fluid (AF) and placenta. When ingested, it enhances opioid-mediated pain relief. Our laboratory has shown that ingestion of AF specifically enhances the hypoalgesia associated with δ-opioid receptor activation in the brain. The specific biochemical compound in AF responsible for the enhancement of δ-opioid activity is of great interest as an analgesic adjunct for pain but is unknown at this time. Research efforts to isolate and characterize this biochemical compound are hampered by the lack of an algesiometric assay that allows repeated measurement of pain threshold and repeated exposure to δ-opioid receptor activation. The cold water tail-flick assay (CWTF) may be a sensitive and reliable pain threshold test of (a) all species of opioids that is (b) not subject to repeated-testing effects. Therefore the CWTF test is potentially ideal for the study of δ opioid systems in a repeated measures design. Here, we confirm these attributes of the CWTF test, and determined that (a) there are no repeated-exposure effects associated with the CWTF assay; (b) there are no repeated-exposure effects associated with repeated central injections of DPDPE ([D-Pen2,D-Pen5]-Enkephalin, a selective δ-opioid agonist) as measured by the CWTF assay; and (c) ingestion of AF in conjunction with a central injection of DPDPE produced the same hypoalgesic enhancement as previously found using another assay.

Long-term reduction in ventral tegmental area dopamine neuron population activity following repeated stimulant or ethanol treatment.

BACKGROUND: Drugs of abuse exert profound effects on the mesolimbic/mesocortical dopaminergic (DA) systems. Few studies have investigated the long-term adaptations in ventral tegmental area (VTA) DA neuron activity after repeated exposure to drugs of abuse. We investigated changes in the electrical activity of VTA DA neurons after cessation from treatment with several stimulants and ethanol. METHODS: Adult rats were treated with stimulants (amphetamine: 2 mg/kg per day, 5 days/week, 2 weeks; cocaine: 15 mg/kg per day, 5 days/week, 2 weeks; nicotine: .5 mg/kg per day, 5 days; ethanol: 10 g/kg per day, 3 weeks) and the single-unit activity of VTA DA neurons was studied in vivo 3 to 6 weeks later. RESULTS: Stimulant and ethanol treatment decreased basal VTA DA neuron population activity but not firing rate or firing pattern. This effect was reversed by acute apomorphine, suggesting that the underlying mechanism for reduced population activity was depolarization inactivation. Anesthesia did not confound this result, as similar effects were observed in amphetamine-treated rats recorded in a conscious preparation. CONCLUSIONS: Reduced basal VTA DA neuron population activity presumably due to depolarization inactivation is a common and long-term neuroadaptation to repeated treatment with stimulants and ethanol. This change in VTA DA neuron activity could underlie the persistent nature of addiction-associated behaviors.

Acute Depletion of D2 Receptors from the Rat Substantia Nigra Alters Dopamine Kinetics in the Dorsal Striatum and Drug Responsivity.

Recent studies have used conditional knockout mice to selectively delete the D2 autoreceptor; however, these approaches result in global deletion of D2 autoreceptors early in development. The present study takes a different approach using RNA interference (RNAi) to knockdown the expression of the D2 receptors (D2R) in the substantia nigra (SN), including dopaminergic neurons, which project primarily to the dorsal striatum (dStr) in adult rats. This approach restricts the knockdown primarily to nigrostriatal pathways, leaving mesolimbic D2 autoreceptors intact. Analyses of dopamine (DA) kinetics in the dStr reveal a decrease in DA transporter (DAT) function in the knockdown rats, an effect not observed in D2 autoreceptor knockout mouse models. SN D2 knockdown rats exhibit a behavioral phenotype characterized by persistent enhancement of locomotor activity in a familiar open field, reduced locomotor responsiveness to high doses of cocaine and the ability to overcome haloperidol-induced immobility on the bar test. Together these results demonstrate that presynaptic D2R can be depleted from specific neuronal populations and implicates nigrostriatal D2R in different behavioral responses to psychotropic drugs.

Analgesic efficacy of orally administered buprenorphine in rats: methodologic considerations.

Buprenorphine has been widely recommended for treatment of pain in rodents. We have previously documented that the recommended postoperative oral dose of buprenorphine in male Long-Evans rats, 0.5 mg/kg, is not as effective as the recommended parenteral dose of buprenorphine (0.05 mg/kg, s.c.) as an analgesic. In the series of experiments reported here, we compared: the analgesic effect of buprenorphine when prepared in two ways in the laboratory with that of a commercially available injectable solution of buprenorphine; the analgesic effect of buprenorphine in Long-Evans rats with that in Sprague-Dawley rats; and Long-Evans and Sprague-Dawley rats for development of pica, a commonly reported side effect of buprenorphine. We followed the pica experiment with assessment of the effectiveness of buprenorphine in establishing a conditioned flavor aversion. The results indicated that method of preparation did not result in any significant differences in the efficacy of injected buprenorphine. Strain of rat was not associated with a significant difference in the efficacy of buprenorphine. However, a significant strain difference was found in development of pica. Buprenorphine treatment was effective in inducing a conditioned flavor aversion. We concluded that the recommended oral dose of buprenorphine (0.5 mg/kg) is ineffective as an analgesic, and that this was not the result of method of preparation of the buprenorphine or strain of rat used. Furthermore, we concluded that buprenorphine treatment may induce gastrointestinal distress in both strains tested. The results reaffirm our previous conclusion that oral administration of buprenorphine at 0.5 mg/kg, despite the general recommendation, is not a reasonable treatment for postsurgical pain in rats.

Low doses of risperidone and morphine interact to produce more analgesia and greater extrapyramidal effects in rats.

The search for non-narcotic drugs that will enhance the analgesic effects of opiates without enhancing their side effects has included the investigation of psychoactive drugs already approved for other uses. Some research has supported an analgesic effect of risperidone (RIS), an atypical neuroleptic. However, the analysis of the analgesic efficacy of RIS alone or as an adjuvant to morphine (MOR) has not considered the production of adverse motor effects that would limit its usefulness as a treatment for pain. We tested whether low doses of RIS would enhance the analgesic action of opiates without inducing untoward motor effects. The analgesia induced by a range of RIS doses (0.1-1.0 mg/kg, SC) was assessed alone and in combination with MOR (5 mg/kg, IP) in male Long-Evans (hooded) rats using two different algesiometric assays: hotplate and tail-flick test. The presence or absence of ptosis, vacuous chewing, and abnormal stationary postures was recorded to evaluate dyskinetic effects. No dose of RIS alone altered pain threshold. However, the highest dose of RIS, 1.0 mg/kg SC, significantly increased the analgesic effects of MOR. Dyskinetic effects of RIS were dose-dependent and enhanced in RIS+MOR treatment. These results do not support the hypothesis that RIS, alone or in combination with MOR, elevates pain threshold without also inducing motor side effects. These findings suggest caution in the use of RIS as either a primary treatment or opiate adjuvant treatment for pain.