Classical Swine Fever Virus vs. Classical Swine Fever Virus: The Superinfection Exclusion Phenomenon in Experimentally Infected Wild Boar

Two groups with three wild boars each were used: Group A (animals 1 to 3) served as the control, and Group B (animals 4 to 6) was postnatally persistently infected with the Cat01 strain of CSFV (primary virus). The animals, six weeks old and clinically healthy, were inoculated with the virulent strain Margarita (secondary virus). For exclusive detection of the Margarita strain, a specific qRT-PCR assay was designed, which proved not to have cross-reactivity with the Cat01 strain. The wild boars persistently infected with CSFV were protected from superinfection by the virulent CSFV Margarita strain, as evidenced by the absence of clinical signs and the absence of Margarita RNA detection in serum, swabs and tissue samples. Additionally, in PBMCs, a well-known target for CSFV viral replication, only the primary infecting virus RNA (Cat01 strain) could be detected, even after the isolation in ST cells, demonstrating SIE at the tissue level in vivo. Furthermore, the data analysis of the Margarita qRT-PCR, by means of calculated ΔCt values, supported that PBMCs from persistently infected animals were substantially protected from superinfection after in vitro inoculation with the Margarita virus strain, while this virus was able to infect naive PBMCs efficiently. In parallel, IFN-α values were undetectable in the sera from animals in Group B after inoculation with the CSFV Margarita strain. Furthermore, these animals were unable to elicit adaptive humoral (no E2-specific or neutralising antibodies) or cellular immune responses (in terms of IFN-γ-producing cells) after inoculation with the second virus. Finally, a sequence analysis could not detect CSFV Margarita RNA in the samples tested from Group B. Our results suggested that the SIE phenomenon might be involved in the evolution and phylogeny of the virus, as well as in CSFV control by vaccination. To the best of our knowledge, this study was one of the first showing efficient suppression of superinfection in animals, especially in the absence of IFN-α, which might be associated with the lack of innate immune mechanisms.     

Experimental acute Babesia caballi infections: I. Red blood cell dynamics

Hematological determinations were made on blood samples from six ponies acutely infected with two dosage levels of Babesia caballi (Group 1: divided into two subgroups of three ponies each). Similar determinations were made on blood samples from three premunized ponies given challenge inoculations (Group 2), and three equidae given uninfected red blood cells (Group 3).A trend towards decreases in RBC counts, hemoglobin concentrations, and hematocrits within one to four days after inoculation (AI) was observed in all groups. However, it was marked only in Group 1. In addition, only in Group 1 was there observed a concerted anemia occurring between Days 7 and 16.Those surviving ponies in Group 1 which developed a higher parasitemia between Days 5 and 6 AI (first parasitemia peak) developed a more severe anemia between Days 7 and 16. Ponies which developed parasitemias higher than 40 × 10³ parasitized cells/mm³ at the first parasitemia peak subsequently died.Free bilirubin in Group 1 animals increased immediately after inoculation, and repeatedly exceeded normal ranges until after Day 20 AI when the RBC counts were rising. Similar changes in free bilirubin did not occur in either Groups 2 or 3. Conjugated bilirubin levels did not exceed normal ranges in any of the experimental animals.Active erythrophagocytosis was evident in histological preparations of lymph node, spleen, liver, and lung from ponies which died. Cytosiderin pigment was present in liver parenchyma, and hematin was scattered throughout lymph nodes and spleen.     

Induction of estrus and ovulation in non-cyclic buffalo (Bubalusbubalis) heifers with progesterone-releasing intravaginal device and pregnant mare serum gonadotrophin and their gonadotrophin profile

A study was undertaken to induce estrus among 15 non-cyclic Murrah buffalo heifers at a relatively early age of 2.5 to 3 yr by progesterone releasing intravaginal device (PRID) application. On Day 13, the PRID was removed and the animals were divided into two groups (A and B). Group B received 1000 IU of pregnant mare serum gonadotrophin (PMSG) intramuscularly (i.m.) immediately after removal of the PRID, whereas Group A was given no further treatment. Circulating gonadotrophin profiles (luteinizing hormone (LH) and follicle stimulating hormone (FSH) were quantified during and after the PRID treatment, as well as during the induced estrous cycle. LH and FSH levels before, during, and after PRID treatment were in the range of 0.5 to 3.0 ng/ml and 10 to 45 ng/ml, respectively, and could be considered basal levels. The peak FSH levels of Group B (PRID + PMSG) during estrus ranged from 69.44 to 337.06 ng/ml, much higher than the levels recorded in Group A (PRID). None of the animals in Group A showed peak LH levels during estrus, whereas two animals in Group B had peak LH levels of 15.84 and 16.93 ng/ml at 0 h and 12 h after detection of estrus. The higher LH and FSH levels obtained in Group B animals compared with Group A animals was possibly due to the superimposed effect of PMSG over PRID. All of the 14 animals exhibited estrus. None of the animals in Group A conceived whereas three out of seven animals in Group B conceived, indicating that PMSG following PRID resulted in ovulatory estrus.     

White spots of the liver in pigs experimentally infected with Ascaris suum

To make clear the relationship between Ascaris suum infection and the appearance of white spot lesions on the surface of the liver in pigs, three groups of pigs were inoculated orally with embryonated A. suum eggs and observed clinicopathologically. Group A of three pigs were inoculated 21 times with 100 eggs each of the nematode, group B of three pigs 4 times with 50,000 eggs each for 10 weeks, and group C of two pigs 2 times with 50,000 eggs each at a one-week interval. All the pigs were sacrificed at the same time 1 week after the final inoculation. Such signs of the nematode infection as dyspnea, coughing and fever appeared in all the pigs of groups B and C seven days after inoculation to continue for several days. In addition, peripheral blood eosinophilia was recorded in these animals 7 or 14 days after inoculation. At autopsy, mesh-worked white spots, some compact and others lymphonodular, were observed on the surface of the liver in all the pigs of the three groups. Main white spots were mesh-worked and lymphonodular in the pigs of group A. They were severe and compact in group B. Therefore, they were rough to the touch. In group C mesh-worked white spots fused with one another and covered the surface of the liver. These white spot lesions observed were morphologically very similar to those found in the field conditions. Complement-fixating antibodies reacting to adult A. suum antigen were detected only in sera from the pigs of group B. Moreover, antibodies involved in the intradermal reaction of immediate type were found in the pigs of groups A and B.     

Clinical and Serological Response after Experimental Inoculation with Babesia Divergens of Newborn Calves with and without Maternal Antibodies

The influence of maternal antibodies on clinical and serological response after experimental inoculation with Babesia divergens of newborn calves was studied. Five calves, born to dams seropositive for B.divergens, (Group 1) had specific maternal antibodies when tested 12 h after their first feeding of colostrum. At that point they were inoculated i.v. with B.divergens infected erythrocytes. Five other calves, born to dams seronegative for B.divergens, (Group 2) had no Babesia specific maternal antibodies when inoculated at the same age. Babesia divergens organisms were demonstrated in blood smears from calves in both groups at some point 5 to 10 days p.i. All calves in both groups had B.divergens specific IgM antibodies at 7 to 17 days p.i. as shown by a modified IF-test. Specific IgG antibodies, transferred by colostrum, were found in all calves of Group 1 before inoculation of B.divergens. The IgG titre of these animals increased by a doubling dilution step at 11–25 days p.i. Among calves of Group 2 specific IgG antibodies were found at first between day 9 and 15 p.i. Both IgM and IgG antibody titres had to be investigated since demonstrated IgG antibodies can originate both from maternally transferred antibodies and from actively produced antibodies after an infection. There was no difference in clinical parameters; parasitaemia, PCV, Hb, and rectal temperature between the groups. This experiment gives evidence that there can be a resistance to bovine babesiosis in newborn calves independent of maternal antibodies.     

Acute and chronic dose of alcohol affect the load carrying capacity of long bone in rats

In the present investigation, the effects of acute and chronic dose of alcohol were evaluated on mechanical properties of long bones of Sprague Dawley rats. In "acute study", 18 animals were divided into three groups containing six animals each, i.e. Group A: control animals, normal saline was given to them intraperitoneally for the period of 5 days; Group B: treated animals, given 20% (v/v) absolute alcohol and Group C: treated animals, given 30% (v/v) absolute alcohol, by same route and time duration. In "chronic study", also, 18 animals were divided into three groups containing six animals each, i.e. Group A: control animals, normal saline was given to them intraperitoneally for the period of 6 weeks; Group B: treated animals, given 20% (v/v) absolute alcohol and Group C: treated animals, given 30% (v/v) absolute alcohol by same route and time duration. A significant increase was observed in bone weight of animals taking 20% alcohol but there was decrease in the same for 30% alcohol in case of acute study. For chronic study, there was a decrease in bone weight for both treated groups. During acute study, breaking strength of bone was increased in case of 20% alcohol administration but a slight decrease was shown in the same for 30% alcohol group as compared to control animals. Breaking strength of long bone in the case of chronic study was decreased in case of both groups taking alcohol, i.e. 20% and 30%. The present document is useful in understanding the functional load carrying capacity of bone during alcoholism.     

Semen changes in boars after experimental infection with porcine reproductive and respiratory syndrome (PRRS) virus

Eleven boars seronegative to porcine reproductive and respiratory syndrome virus (PRRSV) were trained for semen collection: five boars were inoculated intranasally with 6 x 10(6)TCID(50)/ml of PRRSV (Group A); four boars were inoculated intranasally with 6 x 10(4)TCID(50)/ml (Group B); and two boars were used as uninfected control (Group C). Semen samples were collected at 7-d intervals from 49 d prior to experimental inoculation with PRRSV to 70 d after inoculation, and were examined for sperm volume, sperm concentration, sperm morphology, sperm motility and for the presence of PRRSV. The infection in boars was demonstrated by the reisolation of PRRSV from the serum of all inoculated boars. Rectal temperatures and general health of the boars were clinically normal throughout the trial. Differences were observed in the quality of semen collected from boars after experimental infection with PRRSV. This infection induced a significant decrease in sperm motility and in spermatozoa with normal acrosomes. Of the semen samples tested for virus isolation in swine alveolar macrophages PRRSV was only isolated in 1 boar from Group B. The virus was detected in an additional semen sample in Group A by the production of an antibody titer in a biological assay. All attempts to detect PRRSV by RT-PCR in semen samples were unsuccessful. Nevertheless, from our study it is possible to suggest that the PRRSV can occasionally be transmitted in the semen during the initial phase of the disease.     

Induction of parturition in water buffaloes (Bubalus bubalis )

Parturition was induced in ten buffaloes by combining treatments of dexamethasone and vetoestrol, so that they calved about one month before the expected term. Either dexamethasone alone (Group B) or dexamethasone and vetoestrol (Group C) was used. Another five animals served as controls (Group A). Calving was induced in four animals in group B and three animals in group C after two injections of the compounds four days apart. The average time from first injection to calving for these animals was 117.22 hr and 117.66 hr for groups B and C respectively. Induced calves weighed less at birth (P < 0.05) averaging 24.4 and 26.2 kg for groups B and C respectively, than controls (Group A; 30.20 kg). The body weight gain/week among calves was not significantly different (P > 0.05). The service period, number of services/conception and the milk yield of the buffaloes induced to calve was not significantly different (P > 0.05) from their previous records.     

Comparison of pregnancy rates with two estrus synchronization protocols in Italian Mediterranean Buffalo cows

The aim in this study was to compare two estrus synchronization protocols in buffaloes. Animals were divided into two groups: Group A (n=111) received 100 microg GnRH on Day 0, 375 microg PGF(2alpha) on Day 7 and 100 microg GnRH on Day 9 (Ovsynch); Group B (n=117) received an intravaginal drug release device (PRID) containing 1.55 g progesterone and a capsule with 10mg estradiol benzoate for 10 days and were treated with a luteolytic dose of PGF(2alpha) and 1000 IU PMSG at the time of PRID withdrawal. Animals were inseminated twice 18 and 42 h after the second injection of GnRH (Group A) and 60 and 84 h after PGF(2alpha) and PMSG injections (Group B). Progesterone (P(4)) concentrations in milk samples collected 12 and 2 days before treatments were used to determine cyclic and non-cyclic buffaloes, and milk P(4) concentrations 10 days after Artificial insemination (AI) were used as an index of a functional corpus luteum. Cows were palpated per rectum at 40 and 90 days after AI to determine pregnancies. All previously non-cyclic animals in Group B had elevated P(4) (>120 pg/ml milk whey) on Day 10 after AI. Accordingly, a greater (P<0.01) relative percentage of animals with elevated P(4) 10 days after AI were observed in Group B (93.2%) than in Group A (81.1%). However, there was no difference in overall pregnancy rates between the two estrus synchronization protocols (Group A, 36.0%; Group B 28.2%). When only animals with elevated P(4) on Day 10 after AI were considered, pregnancy rate was higher (P<0.05) for animals in Group A (44.4%) than Group B (30.3%). The findings indicated that treatment with PRID can induce ovulation in non-cyclic buffalo cows. However, synchronization of estrus with Ovsynch resulted in a higher pregnancy rate compared with synchronization with PRID, particularly in cyclic buffalo.     

Is slow follicular growth the cause of silent estrus in water buffaloes?

The present experiment was conducted to study the growth profile of the ovulatory follicle in relation to the expression of estrus following administration of PGF(2alpha) to subestrus buffaloes. After detection of a mature corpus luteum by examination per rectum, confirmed by ultrasound scanning, subestrus buffaloes (n=20) were treated (Day 0) with single dose of Dinoprost tromethamin (25 mg, i.m.). Blood samples were collected at 0, 24 and 48 h after treatment for estimation of plasma progesterone concentration. Growth profile of the ovulatory follicle was monitored daily through ultrasound scanning starting from Day 0 until ovulation and the regression profile of CL was monitored at 0, 24 and 48 h of treatment. Estrus was detected by exposure to a fertile buffalo bull three times a day until expression of overt estrus or ovulation. Behavioral estrus was recorded in 14 animals and 6 animals ovulated silently. Sixteen animals including six animals with silent estrus ovulated from the dominant follicle present at treatment (Group A) and remaining four animals ovulated from the dominant follicle of succeeding follicular wave (Group B). The intervals from treatment to estrus (6.5+/-0.25 versus 3.2+/-0.27 days, P<0.001) and treatment to ovulation (7.5+/-0.25 versus 5.4+/-0.46 days, P<0.005) were significantly longer in animals of Group B compared with animals of Group A. Significant differences were observed in growth profile of the ovulatory follicle between animals of Groups A and B with respect to size of the follicle on Day 0 (9.8+/-0.7 versus 5.3+/-0.45 mm, P<0.001), daily growth rate (0.97+/-0.07 versus 1.6+/-0.2 mm/day, P<0.01) and increase in diameter (4.1+/-0.6 versus 7.8+/-0.7 mm, P<0.01). The animals with silent estrus (subgroup A-2) had significantly smaller diameter of the ovulatory follicle on Day 0 (7.7+/-0.4 versus 11.0+/-0.7 mm, P<0.005), its daily growth rate was significantly slower (0.7+/-0.02 versus 1.1+/-0.1 mm/day, P<0.01) and they recorded significantly longer interval from treatment to ovulation (7.3+/-0.56 versus 4.2+/-0.27 days, P<0.001) compared with the animals that showed overt estrus (subgroup A-1). The corpus luteum area (CL area) and plasma progesterone (P(4)) concentration declined continuously from 0 to 48 h after PGF(2alpha) treatment in the animals of both the Groups A and B. Non-significant differences were observed in mean CL area and plasma P(4) concentration at 0, 24 and 48 h post-treatment between animals of Groups A and B and also between animals of subgroups A-1 and A-2. The small size and the slow growth rate of the ovulatory follicle were identified as the possible cause of silent estrus in subestrus buffaloes after PGF(2alpha) treatment.     

Biological characterization of acute infection with ground squirrel hepatitis virus.

Ground squirrel hepatitis virus (GSHV) is a small DNA virus, structurally and antigenically related to the human hepatitis B virus, which occurs naturally among certain wild populations of ground squirrels (P. L. Marion et al., Proc. Natl. Acad. Sci. U.S.A. 77:2941-2945, 1980). Serum from naturally infected animals was used to transmit GSHV in the laboratory by parenteral inoculation of susceptible squirrels. Sixty percent of recipient animals developed viral surface antigenemia after a latent period of 2 to 3 months; three of these animals have remained viremic for over 9 months. Like hepatitis B virus, GSHV demonstrates marked hepatotropism, with viral DNA detected in significant quantities only in the liver, where an average of 6 X 10(2) to 6 X 10(3) viral DNA molecules per cell were found by molecular hybridization. However, histological signs of liver injury after acute infection are minimal. In contrast to infection of its natural host, parenteral administration of GSHV to rats, mice, guinea pigs, and hamsters did not result in demonstrable antigenemia, suggesting that the host range of GSHV, like that of hepatitis B virus, is narrow.     

Comparative experimental infection of Lacazia loboi in BALB/c and B10.A mice

Both hind foot pads of BALB/c and B10.A mice strains, were inoculated with a fungal suspension of Lacazia loboi obtained from a Jorge Lobo's disease patient. The suspension had 9 x 105 cells/ml and its viability index was 45%. The animals were sacrificed at different time periods varying from 24 h to 18 months after inoculation. The BALB/c mice developed an extensive granulomatous infiltrate, similar to the disease in humans, that progressively evolved. The number of fungal elements also increased as the disease progressed, and after the seventh month of inoculation, macroscopic changes of the foot pads were evident. Although the B10.A mice developed an exuberant granulomatous infiltrate, macroscopic changes were not detected. The number of fungal cells in the infected tissues increased in number, but they were lower then the numbers found in the BALB/c strain. The viability indexes were also lower for the B10.A strain. Considering the histopathological findings, the presence of macroscopic changes and the great amount of fungal cells in the infected tissues, the authors concluded that the BALB/c mice strain was more susceptible to L. loboi infection than the B10.A strain.     

Persistent infection of chimpanzees with human immunodeficiency virus: serological responses and properties of reisolated viruses.

Persistent infection by human immunodeficiency virus (HIV-1) in the chimpanzee may be valuable for immunopathologic and potential vaccine evaluation. Two HIV strains, the tissue culture-derived human T-cell lymphotropic virus type IIIB (HTLV-IIIB) and in vivo serially passaged lymphadenopathy-associated virus type 1 (LAV-1), were injected intravenously into chimpanzees. Two animals received HTLV-IIIB as either virus-infected H9 cells or cell-free virus. A third animal received chimpanzee-passaged LAV-1. Evaluation of their sera for virus-specific serologic changes, including neutralizations, was done during a 2-year period. During this period all animals had persistently high titers of antibodies to viral core and envelope antigens. All three animals developed a progressively increasing type-specific neutralizing LAV-1 versus HTLV-IIIB antibody titer during the 2-year observation period which broadened in specificity to include HTLV-HIRF, HTLV-IIIMN, and HTLV-IIICC after 6 to 12 months. The antibody titers against both viruses were still increasing by 2 years after experimental virus inoculation. Sera from all animals were capable of neutralizing both homologously and heterologously reisolated virus from chimpanzees. A slightly more rapid type-specific neutralizing response was noted for the animal receiving HTLV-IIIB-infected cells compared with that for cell-free HTLV-IIIB. Sera from all persistently infected chimpanzees were capable of mediating group-specific antibody-mediated complement-dependent cytolysis of HIV-infected cells derived from all isolates tested. Viruses reisolated from all three animals at 20 months after inoculation revealed very similar peptide maps of their respective envelope gp120s, as determined by two-dimensional chymotrypsin oligopeptide analysis. One peptide, however, from the original HTLV-IIIB-inoculated virus was deleted in viruses from all three animals, and in addition, we noted the appearance of a new or modified peptide which was common to LAV-1 as well as to HTLV-IIIB reisolated from infected chimpanzees. It thus appears that a group-specific neutralizing antibody response as well as a group-specific cytotoxic response can develop in chimpanzees after an inoculation of a single HIV variant. This finding suggests that a common, less immunodominant determinant(s) is present on a single HIV strain which could induce group-specific antibodies during viral infection and replication. The identification of this group-specific epitope and the induction of analogous immunity may be relevant to vaccine development against human acquired immunodeficiency syndrome.     

The effect of shearing procedures on blood levels of growth hormone, cortisol and other stress haematochemical parameters in Sarda sheep

The aim of this research was to investigate how growth hormone (GH) cortisol and some haematochemical parameters could be modified by the stress caused by the stages of shearing in Sarda breed sheep. Five groups of 10 sheep each were formed. Group A, only separated from the flock; Group B, only tied; Group C, both tied and shorn (animals in these three groups were ewe lambs shorn for the first time); Group D, adult females both tied and shorn; and Group E, adult entire males both tied and shorn (animals in these two groups had been shorn previously). Five blood samples were taken from each animal: the day before treatment (first sample); at the start of the treatment (second sample); in the middle of shearing for Groups C, D and E, 10 min after separation in Group A and 10 min after tying in Group B (third sample); at the end of treatment (fourth sample); and on the day after treatment (fifth sample). Plasma GH levels showed a decrease (P < 0.01) in Groups A, B, C and D during treatment (third and fourth samples), while Group E only at the end of shearing (fourth sample). In the third sample, the highest GH levels were recorded for Group E (P < 0.05), while it was recorded in the fourth sample for Groups A and E (P < 0.05). Cortisol levels showed a clear increase (P < 0.01) in all groups during treatment, but Group A showed a decrease in the fourth sample in comparison to the third sample. Males in the second, third and fourth sample and Group A only in the fourth sample showed lower cortisol levels when compared with the other groups (P < 0.05). Plasma glucose levels showed an increase (P < 0.01) in all groups during treatment but Groups B, C and E showed the highest values (P < 0.05). Magnesium (Mg) showed an increase in all groups in the third and fourth sample, while sodium (Na), in the same samples, only in Groups B, C and D. Potassium (K) values showed a significant decrease (P < 0.05) only in Groups C and D at the end of shearing. These results show that GH secretion is influenced by all the stress procedure: separation, tying and shearing. Shearing, even if necessary for animals, causes a significant change of the blood parameters involved in the stress response.     

Morphological and functional characteristics of the dominant follicle and corpus luteum in cattle and their influence on ovarian function

Predicting the functional activity of a dominant follicle (DF) and corpus luteum (CL) might be important before starting a superovulation regime or a synchronization program. The DF and CL were characterized morphologically by using ultrasonography and were characterized functionally by estimating the estradiol-17beta/progesterone (E2/P4) ratio. Their influence on ovarian function was estimated through their ability to ovulate at different stages of development in response to PGF2alpha-application. A total of 47 Holstein Friesian (35 cows and 12 heifers) were used in two experiments. In Experiment 1, 25 animals were examined by daily transrectal palpation and ultrasonography to follow the morphological development of the DF. The status of the DF was categorized into 3 groups (A1, B1, C1). The A1 group (n=7) contained animals with DF in the growing phase or in early static growth phase for less than 3 days. Group B1 (n=13) included animals with DF in static growth phase for 3 to 4 days, while Group C1 (n=5) comprised animals with DF keeping a plateau for more than 4 days or animals with DF in the regression phase. The DF were aspirated transvaginally and the follicular fluid (FF) was analyzed for E2 and P4. In Experiment 2, 22 animals were included. As in Experiment 1, the animals were classified into three groups (A2, n=10; B2, n=5; C2, n=7). They were treated by a single dose of PGF2alpha (25 mg, i.m.) between Days 8 and 12 of the cycle. Results showed that luteolyses occurred in all animals. The DF, which were in growing or in early static growth phase < 3 days were always E2-dominant (E2 > P4) and ovulated after PGF2alpha-application in 6/8 of cases and persisted in 2 (Group A2). The DF persisting > 4 days or that had been in regression were always P4-dominant. This type of DF regressed after PGF2alpha-application (Group C2). The DF in early static growth phase for 3 to 4 days in 5/13 cases were E2-dominant and in 8/13 cases were P4-dominant. This type of DF ovulated in 3/5 cases and regressed in 2/5 cases after PGF2alpha-application (Group B2). These results suggest that the DF is morphologically and functionally defined as long as the DF is in the growing or early static growth phase (A1, A2) for at least 2 days or if the DF is in regression (C1, C2). However, when the DF is in the static growth phase for 3 or 4 days (B1, B2), their morphological and functional characteristics are different. The CL controlls ovulation in the A and C groups and plays an abettor's roll in the B-group.     

Plasma hormone profliles and fertility in ewe lambs given progestagen supplementation after mating

Clun Forest ewe lambs (n = 124) were used to investigate the effects of post-mating progestagen supplementation on fertility. The animals were assigned to 1 of 3 three treatments: Group A (n = 41) served as the controls, Group B (n = 42) received 3 weekly injections of 6 mg of medroxyprogesterone acetate (MAP), while Group C (n = 41) was treated with intravaginal sponge containing 60 mg of MAP; all treatments were administered from Day 5 to Day 26 post mating. Supplementation did not increase the percentage of animals pregnant or those lambing: Group A, 72.2 and 66.6%; Group B, 57.5 and 50.0%; and Group C, 67.5 and 60.0%, respectively. Furthermore, there was no effect of supplementation on plasma progesterone, prolactin, cortisol, growth hormone, insulin, or glucose concentrations (P>0.05). However, pre- and post- mating hormone profiles differed significantly between the animals that lambed or aborted and the animals which were found to be barren at lambing. In the barren animals, progesterone concentrations were lower 4 days before and 9 to 33 days after mating (P<0.01), while overall prolactin concentrations were higher throughout the trial (P<0.01). But there was no difference between barren and fertile lambs in cortisol, growth hormone, insulin or glucose concentrations (P>0.05). These results indicate that progestagen supplementation does not increase the reproductive performance of ewe lambs. However, infertility is associated with reduced luteal function and increased prolactin concentration before and after mating.     

SPERMATOGONIAL CHALONE(S): EFFECT ON THE PHASES OF THE CELL CYCLE OF TYPE A SPERMATOGONIA IN THE RAT

Adult rats with X-irradiated testes were used to analyze the effect of the spermatogonial chalone(s) on the phases of the cell cycle of type A spermatogonia. Twelve days after irradiation, the animals were used in two experiments designed to test the existence of hypothetical G₂ and S phase chalones. For the G₂ assay, rats injected twice with testicular extract (Group I), liver extract (Group II) or physiological saline (Group III) were killed 10 hr after the initial injection. Mitoses of type A, Intermediate and type B spermatogonia were counted in whole mounts of dissected seminiferous tubules. To test for an S phase inhibitor, two groups of rats were given multiple injections of either testicular extract (Group IV) or saline solution (Group V). Twenty-two hr after the first injection they were injected with [³H]thymidine and killed 2 hr later. Silver grains over labelled type A nuclei were counted in radioautographed sections of testes from these animals. The average grain counts were identical in Groups IV and V, indicating that the testicular extract did not affect type A spermatogonia during the S phase. Counts of type A mitoses in Groups I, II and III revealed that in the animals injected with the testicular extract (Group I) the number of divisions was 50% lower than in the control groups (Groups II and III). In contrast, mitotic activity of differentiating spermatogonia (In + B) was similar in all three groups of animals. This result is attributed to a testicular chalone which specifically inhibits type A spermatogonia during the G₂ phase of the cell cycle. Indirect evidence for a G₁ spermatogonial chalone is also presented, as a result of an analysis of published data (Clermont & Mauger, 1974).     

Antiviral antibodies are necessary for control of simian immunodeficiency virus replication

To better define the role of B cells in the control of pathogenic simian immunodeficiency virus (SIV) replication, six rhesus monkeys were depleted of B cells by intravenous infusion of rituximab (anti-CD20) 28 days and 7 days before intravaginal SIVmac239 inoculation and every 21 days thereafter until AIDS developed. Although the blood and tissues were similarly depleted of B cells, anti-SIV immunoglobulin G (IgG) antibody responses were completely blocked in only three of the six animals. In all six animals, levels of viral RNA (vRNA) in plasma peaked at 2 weeks and declined by 4 weeks postinoculation (PI). However, the three animals prevented from making an anti-SIV antibody response had significantly higher plasma vRNA levels through 12 weeks PI (P = 0.012). The remaining three B-cell-depleted animals made moderate anti-SIV IgG antibody responses, maintained moderate plasma SIV loads, and showed an expected rate of disease progression, surviving to 24 weeks PI without developing AIDS. In contrast, all three of the B-cell-depleted animals prevented from making anti-SIV IgG responses developed AIDS by 16 weeks PI (P = 0.0001). These observations indicate that antiviral antibody responses are critical in maintaining effective control of SIV replication at early time points postinfection.