**3. Results**

In 238 group independent blood samples (67.42%) responded to only one serovar with Hardjoprajitino prevalence. In 29 samples (8.51%) Pomona, in 41 (11.6%) Pyrogenes and Wolffi in 18 (5.09%). When two serovars were found, there was again a predominance of the Harjoprajitino serovar in 96 samples (41.9%), Pyrogenes in 53 (23.1%), Pomona in 20 (8.73%), Wolffi in 16 (6.98%), Hardjobovis in 18 (7.8%) and Guaricura in 13 (5.67%). For three serovars, Hardjoprajitino predominated in 25 samples (25.7%), Pyrogenes in 21 (21.6%), Wolffi in 15 (15.40%), Pomona in 12 (12.37%), Copenhageni in 9 (9.27%), Guaricura in 6 (6.18%) and Hardjobovis in 5 (5.15%). For four serovars, Hardjoprajitino was again prevalent in 18 (22.7%) of the samples, Pyrogenes in 15 (18.9%), Pomona and Wolffi in 9 each (11.3%), Copenhageni in 11 (13.9%), Hardjobovis in 7 (8.86%), Guaricura in 5 (6.32%) and CTG in 3 with (3.79%). For five serovars, Hardjoprajitino predominated in 3 samples (25%), each of Pyrogenes, Copenhageni and Pomona in 2 (16.6%), each of Wolffi, Hardjobovis and Guaricura in 1 sample (8.3%). The most frequent serovars were Hardjoprajitino, Pyrogenes, Pomona and Wolffi. These were also prevalent as co-agglutinants for serovars Copenhageni, Guaricura and Hadjobovis with respect to the same serogroup. The large number of serovars with cross-reactions or co-agglutination is noteworthy.

**Figure 1** refers to the 39 animals seroreactive at the first collection (G-2) which remained in the study and shared the same environment with those in G-1.

**Figure 2** summarizes the pregnancy results and the positive percentage of serovars at each moment, also in G-2. Pregnancy rates decreases can be observed at moment 3 with 76.9%, moment 4 with 74.3%, moment 5 with 76.9% and moment 8 with 79.4% of pregnancy.

### **Figure 1.**

*Distribution of results at the beginning of the study (moments 1, 2 and 3) for the G-2 group, according to milk production and pregnancy. Praj = Hardjoprajitino, Po = Pomona, wo = Wolffi, Pyr = Pyrogenes, H=Hardjo, Co = Copenhageni, Ict = Icterohaemorrhagiae, Ctg = Ctg, Gua = Guaricura, min = mini.*

### **Figure 2.**

*Dynamics of the result of pregnancy rates in the G-2 group, at times (Mo 1) to (Mo 9). G-2 milk production in liters of milk can be seen for different moments in* **Figure 3***.*

**Table 1** summarizes the results respective of the number of animals, pregnancy in percentage and production in liters of milk at time 2 in G-2.

Serovar Hardjoprajitino was detected at the moment 6 in 18 animals (46.1%), Pyrogenes in 13 (33.3%), Pomona in 9 animals (23%) and Hardjo in 6 animals (15.3%), when decreases in milk production and fertility were observed for infection by the Hardjo serovar, G-2.

**Figure 4** shows a serovar Hardjoprajitino participation of 69.2% in moment 8, 64.1% in moment 9 and 41% in moment 7, all in G-2. Variation among seroprevalence percentages at those moments can be seen for the various serovars.

Moment 9 (**Figure 4**) had a higher seroprevalence of serovars Hardjoprajitino in 25 animals (64.1%), Pyrogenes in 17 (43.5%), Guaricura in 7 (17.9%), Wolffi in 4 (10.2%), Copenhageni in 3 (7.6%), Pomona in 2 (5.1%), Hardjo in 1 (2.5%) in G-2.

**Figure 5** shows a decrease of pregnancy rates in G-1 at moment 5 which may be related to positivity for serovar Hardjoprajitino. On the other hand, **Figure 6** illustrates G-1 milk production at different times, showing a decrease in milk production at moments 5, 7, 8 and 9 which may be related to calving times. As a matter *Serological Monitoring for* Leptospira *Spp. and Monitoring of Productive and Reproductive… DOI: http://dx.doi.org/10.5772/intechopen.98983*

**Figure 3.**

*Dynamics of milk production in liters, in the group of seroreagent animals (G-2), from Mo 1 to Mo 9.*


### **Table 1.**

*Productivity of seroreagent animals (G-2), pregnancy and production in liters of milk, at time 2. Results expressed as a percentage.*

### **Figure 4.**

*Kinetics (dynamics) of G-2 antibody titers expression, compared to moments 7, 8 and 9 of observation. Praj = Hardjoprajitino, Po = Pomona, wo = Wolffi, Pyr = Pyrogenes, H = Hardjo, Co = Copenhageni, Ict = Icterohaemorrhagiae, Ctg = Ctg, Gua = Guaricura, min = mini.*

of fact, despite attempts to start the experiment with groups as homogeneous as possible, delivery times varied and some animals possibly found themselves in more advanced lactation stages of lactation thus interfering with the group's overall production.

**Table 2** shows all the serovars as percentages, at moment 2 for G-1. The pregnancy rate was 87.7% and the milk production 1,963.8 liters. Comparing these

### **Figure 5.**

*Dynamics of the pregnancy rate in the G-1 group, from the initial moment Mo 1 to Mo 9.*

### **Figure 6.**

*Dynamics of milk production in liters in the G-1 group, from the initial Mo to Mo 9.*


### **Table 2.**

*Productivity of seroreagent animals (G-1), pregnancy and production in liters of milk, at time 2. Results expressed as a percentage.*

pregnancy rates and milk production with those for G-2, both pregnancy rate and milk production is higher for G-1 at the study onset probably due to lower infection rates of serovars such as Hardjoprajitino in G-1.

At moment 3, G-1, serovar Hardjoprajitino was found in 21 animals (42.8%), Pyrogenes in 12 (24.4%), Pomona in 9 (18.3%), Wolffi in 7 (14.2%), Copenhageni *Serological Monitoring for* Leptospira *Spp. and Monitoring of Productive and Reproductive… DOI: http://dx.doi.org/10.5772/intechopen.98983*

in 5 (10.2%) and Icterohaemorrhagiae in 2 (4.08%) with a pregnancy rate of 89.7% and milk production of 1,906.8 liters. Hardjoprajitino remained the most frequent serovar with a slight increase when compared to Mo 2. As mentioned for Mo 2, pregnancy rates and milk production were also higher when compared to the earlier moments, for G-2, where serological response with variable antibody titers for one or more leptospiral serovars were present at study onset. Such observation reinforces the importance especially of the serovar Hardjoprajitino to the productive and reproductive aspects of dairy cattle, the focus of the present study.

**Figure 7** illustrates the dynamics of antibodies titres regarding the Hardjoprajitino serovar with 59.1%, 55.1% and 46.9% positivity thus confirming the relevance of this serovar for cattle. The participation of serovar Pyrogenes among the serovars that stand out at different times is noteworthy.

At moment 4 for G-1, serovar Hardjoprajitino was obtained in 29 animals (59.1%), Pyrogenes in 5 (10.2%), Pomona in 4 (8.16%), Hardjo in 4 (8.16%), Wolffi in 3 (6.12%), Guaricura in 2 (4.08%), Copenhageni in 2 (4.08%), Castellonis in 1 (2.04%). The pregnancy rate was 89.7%, and milk production 1,872 liters of milk. There was a slight decrease in milk production but the pregnancy rate was the same as for the previous month (Mo 3).

### **Figure 7.**

*Kinetics (dynamics) of G-1 antibody titer expression at moments 4, 5 and 6 of observation. Praj = Hardjoprajitino, Po = Pomona, wo = Wolffi, Pyr = Pyrogenes, H = Hardjo, Co = Copenhageni, Ict = Icterohaemorrhagiae, Ctg = Ctg, Gua = Guaricura, min = mini, Ca = Castellonis.*


### **Table 3.**

*Productivity of seroreagent animals (G-1), pregnancy and production in liters of milk, at time 5. Results expressed as a percentage.*

**Table 3** summarizes the G-1 results for moment 5. The pregnancy rate was 75.5%, and milk production volume 1,738 liters. There was a decrease in productive and reproductive indexes with a decrease in pregnancy rate and milk production when compared to previous moments.

Reagent serovars in G-1, moment 6, were Hardjoprajitino in 23 animals (46.9%), Pyrogenes in 8 (16.3%), Pomona in 8 (16.3%), Wolffi in 6 (12.24%), Hardjo in 3 (6.12%), and Copenhageni in 1 (2.04%), with a pregnancy rate of 83.6% and a milk productionn volume of 1,840 liters of milk. There was a decrease in the seroprevalence of Hardjoprajitino from 55.1% to 46.9% and consequently increases in pregnancy rate from 75.5% to 83.6% and in milk production from 1,737.8 to 1,840 liters of milk.

**Figure 8** illustrates the dynamics of the response to serovars, in Mo 7, 8 and 9. There are differences among the various moments, seroprevalence oscillating, which is possible since the animals shared the same environment exposing themselves to animal-maintained and environment serovars.

At moment 7 in G-1, serovar Hardjoprajitino can be observed in 7 animals (14.2%), Pyrogenes in 8 (16.3%), Wolffi in 6 (12.2%), Copenhageni in 3 (6.12%), Pomona in 1 (2.04%), with a pregnancy rate of 87.7%, and milk production of 1,738 liters of milk. Despite the lower response to serovar Hardjoprajitino, there was an increase in the pregnancy rate but a decrease in milk production in comparison to moment 6. At this moment there was no response to serovar Hadjobovis. For this same group, in moment 8, serovar Hardjoprajitino was detected in 22 (44.8%), Pyrogenes in 9 (18.3%), both Guaricura and Hardjo in 6 (12.24%), Ctg in 5 (10.2%), Wolffi, Pomona and Icterohaemorrhagiae in 1 (2.04%) each. The pregnancy rate was 89.7% and milk production volume 1,620 liters of milk. There was an increase in the response to serovar Hardjoprajitino, from 16.3% to 44.8% and the serovar Hardjobovis, not found at moment 7, appears in 12.24% which must have influenced the decrease in milk production from 1,738 to 1,620 liters, despite pregnancy rate showing a slight increase from 87.79% to 89.7%.

**Table 4** summarizes the most frequent serovars for moment 9, with a pregnancy rate of 81.6% and a milk production volume of 1,519 liters. A decrease in the response to serovar Hardjoprajitino as well as a lack of response to serovar Hardjobovis were observed. There was also a decrease in pregnancy rate and milk production despite the lower response to the serovar Hadjoprajitino, a reduction from 44.8% to 24.4% at moments 8 and 9, and the non-response to serovar Hadjobovis at that moment. Regarding the decrease in milk production also seen in G-1, it should be noted that many animals might have been in an advanced stage,

### **Figure 8.**

*Kinetics (dynamics) of G-1 antibody titer expression at moments 7, 8 and 9 of observation. Praj = Hardjoprajitino, Po = Pomona, wo = Wolffi, Pyr = Pyrogenes, H = Hardjo, Co = Copenhageni, Ict = Icterohaemorrhagiae, Ctg = Ctg, Gua = Guaricura, min = mini, Ca = Castellonis.*

*Serological Monitoring for* Leptospira *Spp. and Monitoring of Productive and Reproductive… DOI: http://dx.doi.org/10.5772/intechopen.98983*


### **Table 4.**

*Productivity of seroreagent animals (G-1), pregnancy and production in liters of milk, at the moment 9. Results expressed as a percentage.*

### **Figure 9.**

*Average percentage of animals in the G-1, at moments 1 to 9 of observation. Praj = Hardjoprajitino, Pyr = Pyrogenes, Po = Pomona, wo = Wolffi, H = Hardjo, Co = Copenhageni, Ict = Icterohaemorrhagiae, Ctg = Ctg.*

### **Figure 10.**

*Average percentage of animals in the G-2, at moments 1 to 9 of observation. Praj = Hardjoprajitino, Pyr = Pyrogenes, Po = Pomona, wo = Wolffi, H = Hardjo, Co = Copenhageni, Ict = Icterohaemorrhagiae, Ctg = Ctg.*

near the end of lactation with a consequential decrease in milk production, which was also observed in G-2.

**Figure 9** shows the percentage of response to serovars from Mo 1 to Mo 9 with greater prevalence of serovars Hardjoprajitino, Pyrogenes and Pomona. Regarding the 49 animals in group G-1, **Figure 10** shows the average prevalence of serovars, the decrease in productivity in G-2 at moments 4, 5, 6, 7 and 9 and in G-1 at moments 5 and 9. There was a decrease in the pregnancy rate at moments 3, 4, 5, 6, 8 in group G-2 and at moments 5 and 9 in group G-1.
