Sequential testicular and epididymal damage in Zebu bulls experimentally infected with Trypanosoma vivax.

Six Zebu bulls aged between 31 and 34 months exhibiting good libido were used to study sequential testicular and epididymal damage in Trypanosoma vivax infection. Three bulls were infected with T. vivax, while the other three served as controls. All infected bulls became parasitaemic by day 5 post-infection and developed clinical trypanosomosis with rapidly developing anaemia. Representative bulls, one from each of the infected and control groups, were sacrificed on days 14, 28 and 56 post-infection. Testes and epididymides from these animals were studied histopathologically after processing and staining with haematoxylin and eosin (H and E). Testicular degeneration developed in all the infected bulls characterized by depletion of spermatogenic cells and destruction of interstitial tissue. The most severe testicular degeneration occurred in the bull that was sacrificed 56 days post-infection. Epididymal sperm reserves were 36%, 4% and 0%, respectively, in infected bulls that were sacrificed on days 14, 28 and 56 post-infection. The 0% epididymal sperm reserve may suggest complete cessation of spermatogenesis. It was concluded from this study that T. vivax infection of Zebu bulls could cause severe testicular and epididymal damage that may result in infertility or even sterility of the affected animals at early infection stages not previously thought.

The effects of trypanosomosis on sperm morphology in Zebu x Friesian crossbred bulls.

Detailed studies of sperm morphological abnormalities were carried out on 12 Zebu x Friesian crossbred bulls used in a study of the effects of trypanosomosis. Four bulls were infected with T. vivax, another four with T. congolense, while four served as controls. The infected bulls developed chronic trypanosomosis. All the bulls initially had very low sperm morphological abnormalities that were within acceptable limits for fertile animals. After infection there was a rapid and progressive increase in all sperm abnormalities. Spermatozoa of infected bulls were highly deformed with multiple morphological defects. Mean percentage pre-infection baseline values prior to infection for acrosomal, sperm-head, detached heads, proximal cytoplasmic droplets, distal cytoplasmic droplets, sperm-tail, midpiece and total sperm morphological defects ranged between 0.1 +/- 0.1 for acrosomal and 8.3 +/- 3.2 for total morphological abnormalities in the semen of the bulls. All the infected bulls developed sperm morphological abnormalities of more than a mean of 40.0% from the 4th week after infection until the end of the investigation and were considered unfit for breeding. At 7 weeks post-infection (PI) until the end of the study (12 weeks PI), the controls had a mean of less than 5% sperm morphological defects, while the infected bulls had 100%. Mean percentage values of sperm morphological defects throughout the duration of the investigation for control bulls were low and within the normal range for fertile bulls. These values differed significantly (p<0.001) from the elevated values of the infected bulls. The results show that trypanosomosis due to T. vivax or T. congolense infection can render Zebu x Friesian crossbred bulls unfit for breeding within a very short time. The resultant infertility could be of economic importance in trypanosomosis-endemic sub-Saharan Africa where Zebu x Friesian crossbred bulls are kept.

Effects of nutritional supplementation and exposure to bulls on resumption of post-partum ovarian activity in Bunaji (Bos indicus) cattle.

A total of 51 pluriparous post-partum Bunaji (Zebu) cows belonging to agropastoralists were involved in a 2 x 2 factorial experiment for a period of 180 days. The cows were assigned randomly at calving to four treatments: (1) grazing only (no supplementation) and exposure to bull (NSBE); (2) grazing only (no supplementation) and no exposure to bull (NSNE); (3) grazing plus feed supplementation (each cow received 600g of 20.8% crude protein of whole cottonseed supplement per day) and exposure to bull (FSBE); (4) grazing plus feed supplementation and no exposure to bull (FSNE). Cows with an increase in milk progesterone (P(4)) concentration of > or =1 ng/mL from the weekly milk samples were used to analyse the number of days from calving to the time of resumption of ovarian activity. The mean interval from parturition to cyclic ovarian activity for FSBE cows was 95 days, compared to 119 days for the FSNE cows. Intervals to onset of post-partum ovarian activity were 24, 33 and 39 days which were significantly earlier in the FSBE cows, than the FSNE, NSBE and NSNE cows, respectively. Intervals to cyclic activity were 9 and 15 days earlier in FSNE cows than in NSBE and NSNE cows. By 150 days post-partum, 100% and 92% of the cows in the supplemented groups (FSBE and FSNE), had resumed cyclic ovarian activity compared with 75% and 69% for the unsupplemented cows (NSBE and NSNE). It is concluded that nutritional supplementation and exposure to bulls synergistically shortened the length of post-partum anoestrus in zebu cattle. The economic benefits of using exposure to bulls and cottonseed supplementation in this study to enhance early resumption of post-partum ovarian activity of cattle may serve as a management tool in tropical areas where livestock production has some constraints.

Effects of Trypanosoma vivax and Trypanosoma congolense infections on the reaction time and semen characteristics of Zebu (Bunaji) x Friesian crossbred bulls.

The effect of trypanosomosis on reaction time and semen characteristics of 12 Zebu (Bunaji) x Friesian crossbred bulls aged between 3 and 5 years was studied for a duration of 12 weeks. Four of the bulls were infected with Trypanosoma vivax, another four with Trypanosoma congolense and the remaining four bulls served as controls. Rectal temperatures and haematological parameters were monitored twice weekly. The pre-infection mean value of the rectal temperature was 38.3 degrees C, and this rose to a mean of between 40.5 and 41.1 degrees C in the infected animals. Concurrently, the infected animals exhibited signs of anaemia shown by pale mucous membranes and decreased packed cell volume (PCV), weight loss, lethargy, weakness and dullness. The reaction time (ejaculation time) of semen collection significantly increased from a pre-infection mean value of 20.46-25.14 s to a mean of 290.33-301.15 s within 12 weeks post-infection. Semen characteristics deteriorated progressively within the same period in the infected bulls. There were highly significant and drastic decreases in sperm concentration and volume of semen and increases in sperm morphological defects. By the third week, all the infected bulls were unfit for breeding because of very poor semen characteristics. Deterioration, also characterized by oligospermia at 6 weeks post-infection in all bulls which later culminated in azoospermia in two bulls infected with T. vivax and two bulls infected with T. congolense continued to the end of the investigation. The present results indicate that trypanosomosis due to T. vivax and T. congolense infections is very pathogenic and devastating in its effect on the reaction time (ejaculation time) and semen characteristics which resulted in very poor semen quality. The practical implication is infertility and sterility in Zebu x Friesian crossbred bulls in trypanosome endemic areas.

The role of pheromones and biostimulation in animal reproduction.

It is now known that pheromonal communication plays an important role in mammalian behaviour and reproductive processes. Chemical communication with pheromones is one means of transmitting such information. In mammals, signalling and priming pheromones are thought to act either singly or in combination through olfaction, auditory, visual (sight) or tactile stimuli. Pheromones are air-borne chemical substances ("signals") released in the urine or feces of animals or secreted from cutaneous glands that are perceived by the olfactory system and that elicit both behavioural and endocrine responses in conspecifics. Extensive studies in insects, rodents, swine, sheep, goats and cattle have established the importance of pheromones in the strong influence exerted by the male on reproductive activity in the female. There is a pheromone produced by the queen honey bee, which has two functions: inhibition of queen rearing and suppression of oogenesis in workers and in addition attracts drones during nuptial flight. It has also been demonstrated that the urine of male mice, rats, feral species and other wild rodents contains a priming pheromone that is responsible for hastening puberty in the females. Pheromones in the wool, wax and urine of a ram are sufficient to stimulate ewes to ovulate, while the buck has a strong characteristic seasonal odor and a buck jar containing the odor of the buck can be used as an aid in the detection of oestrus in does. The mere presence of the boar at the time of insemination of the sow improves sperm transport and ovulation, while the presence of the vasectomised bull has been reported to hasten the onset of puberty in heifers and also early resumption of ovarian activity in cattle following parturition. The role of pheromones in bovine reproduction is not as clearly defined as in sheep, goats and swine. Pheromones and other allelomimetic cues can exert profound effects on reproductive activity via the hypothalamic system that generates pulses of gonadotropin-releasing hormone. Manipulations of these factors and other pathways linking environmental inputs to reproductive output can lead to developing the concept of "control systems technologies", aimed at controlling reproductive performance. The knowledge acquired on the effectiveness of biostimulation; the factor which conditions it and the biological mechanism which produces it in livestock species, allows its use as a breeding management tool. The understanding of the role of pheromones could be of potential economic importance in addressing some of the problems associated with livestock production in the tropics. The biostimulation technique offers a potentially useful and practical way to improve reproductive efficiency in livestock species in the tropics. The exact nature of the cues and the role of biostimulation in livestock species especially swine, sheep, goats and cattle in developing countries require more attention.

Effects of Trypanosoma vivax on pregnancy of Yankasa sheep and the results of homidum chloride chemotherapy.

Three groups of pregnant Yankasa ewes, made up of six ewes in each group were assigned at random to first, second and third trimester of pregnancy studies. The ewes were experimentally infected with T. vivax to study the effects of the infection on pregnancy and the results of Novidium Chemotherapy. Three pregnant uninfected ewes served as controls. Fourteen days post infection, the ewes in each trimester study, were paired by weight and assigned to two groups of three ewes each. One group was treated with Novidium while the other group remained untreated. Of the three ewes in each group, one ewe was killed humanely at 21 days post infection and another at the end of the trimester period. In the first trimester, a ewe with partial fetal resorption was observed among the untreated ewes. Fetal death in-utero and expulsion of an autolyzed fetus was observed among the treated ewes. In the second trimester, abortion and almost complete fetal resorption were observed among the untreated ewes. Fetal death in-utero and expulsion of an autolyzed fetus was observed among the treated ewes. In the third trimester, abortions were observed among the untreated ewes. Abortion of a live fetus and a case of dystocia were observed among the treated ewes. Ewes in the second and third trimesters of pregnancy were more susceptible to the infection, with ewes in the third trimester being most susceptible, as measured by the number of abortions and death of ewes. Fetuses from the untreated ewes in the three trimesters of pregnancy were lower in body weights, than the fetuses from the treated ewes. The uninfected control ewes carried the pregnancies to term. Novidium chemotherapy at 14 days post infection was not beneficial in ameliorating the pathogenicity of T. vivax infection on pregnancy in Yankasa ewes. T. vivax infection of only 14 days was enough to cause irreversible pathology in Yankasa fetuses evidenced by death of fetuses in-utero, dystocia and abortions irrespective of Novidium chemotherapy.

Effects of bull exposure and body growth on onset of puberty in Bunaji and Friesian x Bunaji heifers.

A total of ninety seven pre-pubertal Bunaji (BJ) and Friesian-Bunaji (FR x BJ) heifers were allotted randomly to two treatments groups for a period of 15 months. The treatment groups consisted of the followings: Mature Bull Exposure (MBE) and No Bull Exposure (NBE). Heifers were body condition scored and their live weights recorded on 28 days consecutive intervals. A heifer tactile stimulation in bull bio-stimulation. was considered to have attained puberty if she displayed oestrus, had a palpable corpus luteum with an associated P4 concentration > 1 ng x mL(-1). The onset of puberty was significantly earlier in MBE heifers (23.1 +/- 0.4 months) than NBE heifers (26.4 +/- 0.4 months). The mean ages at puberty for MBE-BJ, NBE-BJ, MBE-FR x BJ, NBE-FR x BJ were 24.3, 27.8, 22.1 and 25.0 months respectively. More MBE heifers (70.8%) attained puberty between 17 and 24 months of age than NBE heifers (18.3%) and on the same ages, more FR x BJ heifers (62.0%) than BJ heifers (25.5%). The mean live weight of MBE heifers at puberty (224.4 +/- 4.2 kg) was significantly lower than that of the NBE heifers (255.8 +/- 4.2 kg). The FR x BJ heifers attained puberty at a significantly higher live weight (270.2 +/- 4.2 kg) than the BJ heifers (228.6 +/- 4.2 kg). The use of a vasectomised bull especially in some elite farms that rely on artificial insemination services may be an effective management tool that can decrease age at puberty. More work is required to determine the relative contribution of visual, auditory, olfactory, pheromonal and tactile stimulation in bull biostimulation.

Serum progesterone profiles of zebu cattle (Bos indicus) in relationship to conception and repeat breeding after artificial insemination.

Sixty-four zebu cows, comprised mainly of Bunaji cattle aged between 3 and 7 years old, were involved in an intensive artificial insemination programme. The study was conducted during the breeding period (May-October), and cows were inseminated with freshly prepared Friesian semen when they stood to be mounted by the vasectomised bull or herdmates. Fertility was measured by serum progesterone (P(4)) concentrations and pregnancy diagnosis by rectal examination post-insemination. From the records of oestrus detection and P(4) profiles of cattle following breeding, 39 oestrous cycle lengths were classified into short, normal and long cycles. The mean duration for short, normal and long oestrous cycles were 15.6+/-2.0, 21.5+/-1.5 and 29.5+/-2.5 days, respectively. Mean inter-oestrus intervals between the treatment groups were not different. The percentage frequency distribution was 48.7% for normal oestrous cycles. The short oestrous cycle lengths of 11-17 days were observed in 12.8% of the cases, while 38.5% of the cases of oestrus returns had long luteal phases with oestrous cycle lengths of 26-32 days. Out of the 64 cows, 48.4% conceived to the first breeding while 18.8% conceived to the second breeding. Five and nine cows became acyclic after the first and second breeding, respectively, constituting 22. 0%. Cows that displayed irregular oestrous cycles (repeat breeders) constituted 10.9%. Average number of services per conception was 1.3. Serum P(4) is of practical value in monitoring ovarian activity in cattle and in the identification of acyclic and repeat breeder cows. Repeat breeder cows could be investigated by a combination of radioimmunoassay and clinical examination of the ovaries and reproductive tract. In cattle management, it is economical and profitable to diagnose pregnancy early after insemination so that cows which fail to conceive may be rebred.

Fertility in zebu cattle (Bos indicus) after prostaglandin administration and artificial insemination.

A total of 137 cycling zebu cows, each receiving a single dose of prostaglandin PGF(2alpha)were used in an oestrus synchronization programme on three different farms. Of the cows on the three farms, 60.6 and 90.5% showed overt oestrus and luteolysis, respectively. Pregnancy rate to fixed time inseminations following single injection of PGF(2alpha)was 61.4% for farm 1, significantly higher than the values of 45.7 and 46.9% for farms 2 and 3, respectively. The pregnancy rates to second service of rebred cows were 53.3, 50.0 and 50.0% for the three farms, respectively, with no significant differences between each. Fertility classification of the cows based on progesterone (P(4)) concentration showed that 6.6% of cows on the three farms were incorrectly diagnosed as having corpora lutea; 2.9% of them had incomplete luteolysis and 5.1% may have lost their embryos between days 21 and 45 post-insemination. The pregnancy rate was 10% higher in the rainy season than in the dry season. Cows with body condition scores of 3 and 4 had a higher overall pregnancy rates than those with a body condition score of 2. The findings of this study further confirm the luteolytic efficacy of prostaglandin in inducing oestrus in zebu cattle and indicate that the nutritional status of the cows must be satisfactory before embarking on oestrus synchronization programmes.

Age and hourly related changes of serum testosterone and spermiogram of prepubertal bulls fed two levels of nutrition.

Serum testosterone concentrations and the spermiograms of prepuberal bulls fed two levels of protein diets were investigated at 7, 10, 14 and 18 months of age. Scrotal circumference, body condition score and total sperm counts of those animals on a high protein diet were significantly (P < 0.05) higher than those on a low protein diet. However, sperm motility, total dead sperm and abnormal sperm did not differ between the treatment groups (P < 0.05). One bull fed a high protein diet had significantly higher testosterone concentrations (basal and peak) than a bull fed low protein throughout the four sampling periods (P < 0.05). Testosterone concentrations, scrotal circumference, volume of semen, sperm concentration and sperm output of bulls on low and high protein diets increased significantly with age (P < 0.05). Peak testosterone concentrations ranged from 1.1 ng ml-1 at 7 months to a maximum of 5.3 ng ml-1 at 18 months. The 24 h secretory patterns of testosterone were episodic, pulsatile or temporal in nature. The peaks occurred mostly in the morning hours and ranged from one to five in number. Protein intake in prepubertal bulls could have significant influence on spermiogram and testosterone production.

Reproductive disorders caused by animal trypanosomiases: a review.

Pathogenic animal trypanosomes are causative agents of the most common livestock diseases which have an important economic impact on many African countries. These diseases usually cause debilitating symptoms manifested by anemia and cachexia which may result in death. Recent studies show that they cause a wide range of reproductive disorders in animals, including degeneration of the hypothalamus, pituitary glands and gonads with consequent disruptions in the secretions and plasma concentrations of the hormones necessary for normal reproductive processes in both sexes. Reproductive disorders caused in male animals include delayed puberty, loss of libido, severe degenerative changes of the genitalia manifested by the production of very poor quality semen or the cessation of semen production. In female animals trypanosomiases cause severe genital lesions, temporary or permanent anestrus, and abnormal estrous cycles. Additionally, trypanosomal-induced death during pregnancy, abnormal pregnancy, dystocia, abortion, premature birth, low birth weight, stillbirth, transplacental fetal infection, neonatal death and other pathogenic effects on fetuses and offspring have been reported. Early treatment with trypanocides may prevent some of the trypanosomal-induced reproductive disorders and the resolution of mild genital lesions. Trypanosomal-induced reproductive disorders in animals are of significant economic importance, especially in sub-Saharan Africa, where tsetse transmitted trypanosomiases are endemic.

Effects of severe chronic scrotal Dermatophilus congolensis (kirchi) infection on semen characteristics in Zebu/Friesian crossbred bulls and effect of long-acting terramycin chemotherapy.

The semen characteristics of 12 Zebu/Friesian crossbred bulls, aged 2 to 3 years, were studied during a 21-month period. At the 12th month of the study, the commencement of the rainy season, the bulls were infected naturally with Dermatophilus congolensis . Lesions were scattered over the body and limbs, but were particularly pronounced on the scrotum. Monthly treatments with injection of terramycin were begun as soon as lesions were detected and continued until the end of the study. The lesions worsened and became pronounced particularly on the scrotum of all the bulls. Scrotal scab formation caused by infection became prominent at the 14th month of the study. Until that period, the bulls had normal semen characteristics. From the 15th month until the end of the study, there was progressive deterioration of semen characteristics in all the bulls; this was manifested by some or all of the following effects: decreased volume, increased percentage of dead spermatozoa, increased percentage of total sperm morphological abnormalities, decreased percentage of progressively motile spermatozoa, oligospermia and terminal azoospermia. Therefore, severe chronic scrotal dermatophilosis may be a significant cause of infertility or sterility in bulls.

Effect of Trypanosoma vivax infection on semen characteristics of Yankasa rams.

Twelve Yankasa rams aged between 2 1/2 and 3 years with good semen characteristics were used in this 15-week study. Six rams were infected with Trypanosoma vivax, while six served as controls. The infected rams developed chronic trypanosomosis accompanied by fluctuating pyrexia, lethargy, anaemia, scrotal oedema and cachexia. There was a drastic and progressive deterioration in semen quality in all infected rams manifested by a decrease in volume or cessation of semen production, oligozoospermia, a sharp decrease in progressively motile sperm, elevated numbers of dead (eosinophilic) sperm and 100% morphological abnormalities of sperm in most animals. The rams were all deemed unfit for breeding by 3 weeks post-infection. Uninfected rams were healthy and had good semen characteristics throughout the investigation. The results show that rams infected with T. vivax may become infertile within a short interval due to rapid deterioration of semen characteristics and this trypanosome species may be an important causative agent of infertility in endemic areas.

Effect of chemotherapy on elevated ejaculation time and deteriorated semen characteristics consequent to bovine trypanosomiasis.

The effect of the trypanocidal drug Novidium on elevated ejaculation time and deteriorated semen characteristics was studied in Zebu cattle infected with T. vivax and T. congolense. Two groups, comprising six bulls per group, were infected with Trypanosoma vivax or Trypanosoma congolense while three bulls served as controls. Chemotherapy was carried out 12 weeks post-infection on three bulls from each group, leaving three bulls untreated while three bulls served as uninfected controls. Blood samples from treated bulls were all negative for trypanosomes 3 days post-chemotherapy. The animals also had normal body temperature. As the study progressed, clinical signs associated with trypanosomiasis, such as anaemia and cachexia, disappeared gradually in treated bulls. There was some improvement in semen characteristics of some of the bulls at 10 weeks post-chemotherapy with Novidium. However, all bulls infected with T. vivax or T. congolense irrespective of Novidium chemotherapy still had poor semen characteristics manifested by all or some of the following: decreased volume of semen, oligospermia, azoospermia and elevated incidence of spermatozoa morphological abnormalities. They were thus unsuitable for breeding.

Elevation of morphological abnormalities of spermatozoa in the semen of Zebu bulls consequent to Trypanosoma vivax and Trypanosoma congolense infections.

Twenty-four Zebu bulls were used in a 12-wk long study. Eight bulls were infected with T. vivax , eight others with T. congolense and eight bulls served as controls. All the infected bulls developed chronic trypanosomiasis. Mean percentage base-line values prior to infection for acrosomal, sperm-head, detached heads, proximal cytoplasmic droplets, distal cytoplasmic droplets, sperm-tail, midpiece and total sperm morphological abnormalities ranged between 0.1+/-0.1 for acrosomal and 8.7+/-3.4 for total morphological abnormalities in the semen of the bulls. These values were very low and within the range of those for fertile bulls. Following infection, there was a progressive increase in the mean values of all the abnormalities. Peak percentage mean values recorded for total sperm morphological abnormalities in the course of the investigation in the bulls infected with T. vivax and T. congolense and in the controls were 95+/-7.2, 100+/-0 and 7.9+/-5.0, respectively. Mean percentage values throughout the duration of the investigation for control bulls were low and within the normal range for fertile bulls. These values differed (P<0.001) from the elevated values of the infected bulls. The results indicate that trypanosomiasis due to either T. vivax or T. congolense infections can cause a marked increase in morphological abnormalities of spermatozoa which can, in turn reduce the fertility of breeding bulls.

Effect of novidium (homidium chloride) chemotherapy on genital lesions induced by Trypanosoma vivax and Trypanosoma congolense infections in Zebu bulls.

Zebu bulls chronically infected with Trypanosoma vivax and T. congolense were treated at the 12th week post-infection with novidium and slaughtered at different times after treatment to determine histological evidence of healing of the genital lesions. Though trypanosomes disappeared from the blood soon after chemotherapy, there was incomplete resolution of genital lesions even 10-18 weeks later. Where there is severe degeneration of the testes and epididymes chemotherapy may be ineffective in leading to regeneration and therefore infertility problems may persist in bulls with chronic trypanosomiasis which are later subjected to chemotherapy.

Pathological changes in male genitalia of cattle infected with Trypanosoma vivax and Trypanosoma congolense.

Samples for histological studies were taken from the genitalia of 14 bulls (five infected with Trypanosoma vivax, five with T. congolense and four uninfected control animals), slaughtered 12, 22 or 30 weeks post-infection. Infection with Y58 strain of T. vivax and strain 2295 of T. congolense caused various grades of lesions in the male reproductive organs, especially the testes and epididymides. T. congolense produced more severe degenerative changes than T. vivax. It is concluded that in long-standing cases, the result of trypanosome infection is either serious infertility or even sterility.

Comparative haematological changes following Trypanosoma vivax and T. congolense infections in Zebu bulls.

A comparative study of haematological changes subsequent to Trypanosoma vivax and Trypanosoma congolense infections was carried out using 24 Zebu bulls during a period of 12 weeks. Eight bulls were infected with T. vivax, another eight with T. congolense and eight served as controls. Infected bulls developed chronic trypanosomiasis which was characterized by many clinical manifestations including intermittent pyrexia. Elevated rectal temperatures of up to 105 and 106 degrees F were recorded, respectively, in all bulls infected with T. vivax or T. congolense. Mean parasitaemia was higher and more chronic in T. congolense-infected bulls and ranged between means of 0 and 3.06 in all infected bulls. There was a slight and transient drop in packed cell volume (PCV) of T. vivax-infected bulls as against a marked and more chronic drop in T. congolense-infected bulls. Mean PCV of T. vivax- and T. congolense-infected bulls and controls ranged between 28 and 38, 17 and 38, and 31 and 38%, respectively. Haemoglobin (Hb) concentrations also decreased in infected bulls. The decrease was greater and more chronic in the T. congolense-infected bulls. Values ranged between means of 10.47 and 13.84, 5.44 and 14.16, and 10.24 and 14.12 g dl-1 in T. vivax- and T. congolense-infected and control bulls, respectively. Total plasma proteins also decreased in infected bulls; this was more marked in the T. congolense-infected group. Values for T. vivax-infected, T. congolense-infected and control bulls ranged between means of 7.66 and 8.99, 6.26 and 8.81, and 7.94 and 8.78 g dl-1, respectively.2+ that the indigenous T. vivax strains are more